U.S. patent application number 14/357555 was filed with the patent office on 2015-01-22 for evaluation of presence of and vulnerability to atrial fibrillation and other indications using matrix metalloproteinase-based imaging.
This patent application is currently assigned to Lantheus Medical Imaging, Inc.. The applicant listed for this patent is Lantheus Medical Imaging, Inc., Yale University. Invention is credited to Joseph G. Akar, Richard R. Cesati, Stephen B. Haber, Heike S. Radeke, Albert J. Sinusas.
Application Number | 20150023873 14/357555 |
Document ID | / |
Family ID | 48290460 |
Filed Date | 2015-01-22 |
United States Patent
Application |
20150023873 |
Kind Code |
A1 |
Sinusas; Albert J. ; et
al. |
January 22, 2015 |
EVALUATION OF PRESENCE OF AND VULNERABILITY TO ATRIAL FIBRILLATION
AND OTHER INDICATIONS USING MATRIX METALLOPROTEINASE-BASED
IMAGING
Abstract
The invention provides, in some embodiments, methods relating to
assessing increased risk of developing atrial fibrillation (AF),
and/or the likelihood of responding to particular AF therapies
using imaging agents comprising an MMP inhibitor linked to an
imaging moiety. The invention further provides methods for
evaluating the presence of the risk of developing other
cardiovascular conditions and assessing the effectiveness of
treatment or other intervention for such conditions by determining
MMP levels.
Inventors: |
Sinusas; Albert J.;
(Guilford, CT) ; Akar; Joseph G.; (New Haven,
CT) ; Cesati; Richard R.; (Pepperell, MA) ;
Radeke; Heike S.; (South Grafton, MA) ; Haber;
Stephen B.; (Westford, MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Lantheus Medical Imaging, Inc.
Yale University |
North Billerica
New Haven |
MA
CT |
US
US |
|
|
Assignee: |
Lantheus Medical Imaging,
Inc.
North Billerica
MA
Yale University
New Haven
CT
|
Family ID: |
48290460 |
Appl. No.: |
14/357555 |
Filed: |
October 31, 2012 |
PCT Filed: |
October 31, 2012 |
PCT NO: |
PCT/US2012/062863 |
371 Date: |
May 9, 2014 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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|
61558677 |
Nov 11, 2011 |
|
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|
61559587 |
Nov 14, 2011 |
|
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61658531 |
Jun 12, 2012 |
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Current U.S.
Class: |
424/1.69 ;
424/1.65; 424/1.89 |
Current CPC
Class: |
A61K 51/044 20130101;
A61K 51/0478 20130101; A61K 51/0476 20130101; A61K 51/0459
20130101; A61B 2576/023 20130101; A61K 51/0497 20130101; A61K 51/08
20130101 |
Class at
Publication: |
424/1.69 ;
424/1.89; 424/1.65 |
International
Class: |
A61K 51/04 20060101
A61K051/04; A61K 51/08 20060101 A61K051/08 |
Claims
1. A method of evaluating risk of developing atrial fibrillation
comprising administering to a subject an imaging agent comprising a
matrix metalloproteinase (MMP) inhibitor linked to an imaging
moiety, and acquiring a cardiac image of the subject, wherein a
level of the imaging agent in the heart of the subject above
control is indicative of an increased risk of developing atrial
fibrillation.
2. A method for evaluating risk of developing atrial fibrillation
recurrence comprising administering to a subject previously
diagnosed with atrial fibrillation and previously treated for
atrial fibrillation an imaging agent comprising a matrix
metalloproteinase (MMP) inhibitor linked to an imaging moiety, and
acquiring a cardiac image of the subject, wherein a level of
imaging agent in the heart of the subject above control is
indicative of an increased risk of atrial fibrillation
recurrence.
3. The method of claim 2, wherein the atrial fibrillation
recurrence is atrial fibrillation recurrence following
cardioversion therapy.
4. A method for identifying a subject having a history of atrial
fibrillation that is likely to respond to treatment with an
implantable pacer, pharmacological rate control therapy,
pharmacological rhythm control therapy, or ablation therapy
comprising administering to a subject previously diagnosed with
atrial fibrillation an imaging agent comprising a matrix
metalloproteinase (MMP) inhibitor linked to an imaging moiety, and
acquiring a cardiac image of the subject, wherein a level of
imaging agent in the heart of the subject above control identifies
a subject to be treated with an implantable pacer.
5-7. (canceled)
8. The method of claim 4, wherein the subject has experienced one
AF event.
9. The method of claim 4, wherein the subject has experienced
recurrent AF.
10. The method of claim 1, wherein the cardiac image is an atrial
image.
11. The method of claim 10, wherein the cardiac image is a left
atrial image.
12. The method of claim 1, wherein the subject is a human
subject.
13. The method of claim 1, wherein the subject does not manifest
signs associated with myocardial fibrosis.
14. The method of claim 1, wherein the subject does not manifest
signs associated with myocardial remodeling.
15. The method of claim 1, wherein the subject has experienced a
myocardial infarction.
16. The method of claim 1, wherein the imaging agent is RP805.
17. The method of claim 1, wherein the imaging agent is selected
from the group consisting of ##STR00096## wherein F represents an
isotopically-enriched population of .sup.18F, and variants thereof
comprising, instead of F, an isotopically-enriched imaging moiety
selected from the group consisting of .sup.11C, .sup.13N,
.sup.123I, .sup.125I, .sup.99mTc, .sup.95Tc, .sup.111In, .sup.62Cu,
.sup.64Cu, .sup.67Ga, and .sup.68Ga.
18. The method of claim 1, further comprising determining a measure
of myocardial perfusion in the subject.
19. The method of claim 18, wherein determining a measure of
myocardial perfusion in the subject comprises administering to the
subject a myocardial perfusion imaging agent and obtaining a
myocardial perfusion image to determine a measure of myocardial
perfusion.
Description
BACKGROUND OF INVENTION
[0001] Atrial fibrillation (AF) is a disturbance in the rhythmic
beating (or arrhythmia) of the upper chambers of the heart. AF is
the most common sustained cardiac arrhythmia, responsible for
almost 50% of hospitalizations for arrhythmias (Benjamin E J, et
al. 1998; Wyse D G, et al. 2002; Benjamin E J, et al. 1997;
Allessie M A, et al. 2001). Consequently, AF is a significant cause
of morbidity and mortality and treatment of AF has been hampered by
the ineffectiveness of available drugs. Moreover, attempts to
terminate AF with electrical shocks, using a process termed
"cardioversion" works for only about one-half of the patients
during the first 6-12 months. Therefore, a "simple" means to
identify patients in whom cardioversion would effectively terminate
AF would be beneficial in terms of saved time and money. Moreover,
a means to identify patients that may or may not benefit from an
implantable pacer, pharmacological rate and/or rhythm control
therapy and/or ablation would also be beneficial.
SUMMARY OF INVENTION
[0002] The invention is broadly based on the use of matrix
metalloproteinase (MMP) based imaging to detect the presence of
(including early detection of) and/or the risk of developing
certain cardiovascular conditions and to evaluate the efficacy of
therapies and/or other interventions directed towards such
conditions. MMP levels (and thus activity) can be detected using in
vivo medial imaging techniques. In accordance with the invention,
MMP activation is an indicator of tissue (e.g., vascular)
remodeling, and such remodeling is an indicator of certain
conditions or an indicator of developing certain conditions.
[0003] Thus, the invention is premised, in part, on the surprising
finding that MMP levels (and activity) can be detected at early
time periods, including prior to the onset of certain
cardiovascular conditions such as but not limited to myocardial
fibrosis, AF, calcific aortic valve disease (CAVD), and
atherosclerosis. It has not been heretofore known that MMP levels
could be detected at such early time points and that detection at
these early time points correlated with increased risk of, for
example, AF and CAVD. The invention therefore provides, inter alia,
methods for determining increased risk (or likelihood, as the terms
are used interchangeably herein) of developing, for example, AF or
CAVD based on detection and/or quantification of MMP levels, and
more specifically localized MMP levels such as cardiac MMP
levels.
[0004] In one aspect, the invention provides a method for
evaluating the risk of developing AF comprising administering to a
subject an imaging agent comprising a matrix metalloproteinase
(MMP) inhibitor linked to an imaging moiety and obtaining a cardiac
image.
[0005] In some embodiments, the subject has experienced a
cardiovascular insult such as, but not limited to, a myocardial
infarction and/or heart surgery and/or has been diagnosed as having
a cardiovascular disease such as but not limited to coronary artery
disease, heart valve disease, cardiomyopathy, and/or congenital
heart disease. If the subject has experienced a cardiovascular
insult, the method may be performed within days, or within a week,
or within 2 weeks, or within a month, or within 2 months of the
cardiovascular insult. In some embodiments, the method may be
performed within 10 days of the cardiovascular insult. In some
embodiments, the subject does not have a history of AF (i.e., the
subject has not been heretofore diagnosed as having AF), although
the subject may have been diagnosed with other cardiovascular
diseases as stated above. In some embodiments, the subject may have
a familial history of AF and/or the subject may present with
certain risk factors that are associated with an increased risk of
developing AF. Such risk factors include high blood pressure and
chronic lung disease.
[0006] In these and other aspects of the invention, the method may
be performed once on the subject or it may be performed repeatedly
on the subject in order to monitor the subject over a period of
time.
[0007] In another aspect, the invention provides a method for
evaluating the risk of AF recurrence (e.g., following cardioversion
therapy). The method comprises administering to a subject
previously diagnosed with AF and treated with an AF therapy an
imaging agent of the invention, and obtaining an image of the heart
of the subject, in whole or in part. In some embodiments, the
above-normal levels (including in some instances mere presence) of
MMP following cardioversion therapy indicate an increased risk of
AF recurrence (e.g., following cardioversion). In some embodiments,
the AF therapy may be cardioversion, pharmacological rate and/or
rhythm control, an implantable pacer, and the like.
[0008] In another aspect, the invention provides a method for
identifying in a subject having a history of AF the likelihood of
responding to treatment with an implantable pacer. The method
comprises administering to a subject previously diagnosed with AF
an imaging agent of the invention, and obtaining an image of the
heart of the subject, in whole or in part. The above-normal levels
(including in some instances mere presence) of MMP identify the
subject as one to be treated with an implantable pacer.
[0009] In another aspect, the invention provides a method for
identifying in a subject having a history of AF the likelihood of
responding to pharmacological rate control therapy. The method
comprises administering to a subject previously diagnosed with AF
an imaging agent of the invention, and obtaining an image of the
heart of the subject, in whole or in part. The above-normal levels
(including in some instances mere presence) of MMP identify the
subject as one to be treated using pharmacological rate control
therapy. Such therapies include beta-blockers, calcium antagonists,
and digoxin.
[0010] In another aspect, the invention provides a method for
identifying in a subject having a history of AF the likelihood of
responding to pharmacological rhythm control therapy. The method
comprises administering to a subject previously diagnosed with AF
an imaging agent of the invention, and obtaining an image of the
heart of the subject, in whole or in part. The above-normal levels
(including in some instances mere presence) of MMP identify the
subject as one to be treated using pharmacological rhythm control
therapy. Such therapies include beta-blockers, amiodarone, class Ic
agents, and sotalol.
[0011] In another aspect, the invention provides a method for
identifying in a subject having a history of AF the likelihood of
responding to ablation therapy. The method comprises administering
to a subject previously diagnosed with AF an imaging agent of the
invention, and obtaining an image of the heart of the subject, in
whole or in part. The above-normal levels (including in some
instances the mere presence) of MMP identify the subject as one to
be treated using ablation therapy.
[0012] In some embodiments, the subject having a history of AF is a
subject that has experienced an AF event. In some embodiments, the
subject having a history of AF is a subject that has experienced
recurrent AF.
[0013] The image indicates the presence or absence of imaging agent
in the heart of the subject which in turn indicates the level (or
amount of MMP) in the heart of the subject. In some embodiments,
the image indicates the amount of the imaging agent in the heart of
the subject, in whole or in part. In some embodiments, the image
comprises atrial myocardium, including the left atrial myocardium,
of the subject.
[0014] In some embodiments, the MMP inhibitor has an inhibitory
constant K.sub.i of <1000 nM. In some embodiments, the MMP
inhibitor has an inhibitory constant K.sub.i of <100 nM. In some
embodiments, the MMP inhibitor is an inhibitor of one or more
matrix metalloproteinases selected from the group consisting of
MMP-2, MMP-9 and MMP-14.
[0015] In some embodiments, the imaging agent is
##STR00001##
(referred to herein as .sup.99mTc-RP805).
[0016] In some embodiments, the imaging agent is
##STR00002##
(referred to herein as .sup.111In-RP782).
[0017] In some embodiments, the imaging agent of the invention is
selected from the group consisting of:
##STR00003##
wherein F represents an isotopically-enriched population of
.sup.18F, and variants thereof comprising, instead of F, an
isotopically-enriched imaging moiety selected from the group
consisting of .sup.11C, .sup.13N, .sup.123I, .sup.125I, .sup.99mTc,
.sup.95Tc, .sup.121In, .sup.62Cu, .sup.64Cu, .sup.67Ga, and
.sup.68Ga. That is, in some embodiments, F is replaced with an
isotopically-enriched imaging moiety or a chelator associated with
an isotopically-enriched imaging moiety, wherein the imaging moiety
is selected from the group consisting of .sup.11C, .sup.13N,
.sup.123I, .sup.125I, .sup.99mTc, .sup.95Tc, .sup.111In, .sup.62Cu,
.sup.64Cu, .sup.67Ga, and .sup.68Ga.
[0018] Other suitable imaging agents for MMP detection and
quantitation are described herein.
[0019] In some embodiments, the method further comprises
determining myocardial perfusion in the subject. Myocardial
perfusion may be determined by obtaining an image of the heart, in
whole or in part, using a myocardial perfusion imaging agent.
Suitable myocardial perfusion imaging agents include flurpiridaz F
18 injection
(4-chloro-2-(1,1-dimethylethyl)-5-({4-[(2-[.sup.18F]fluoroethoxy)methyl]p-
henyl}methoxy)pyridazin-3(2H)-one), .sup.99mTc sestamibi
(Tc99m[MIBI].sub.6 where MIBI is 2-methoxy isobutyl isonitrile),
.sup.201Thallium, and the like. In some embodiments, myocardial
perfusion is quantitated. In some embodiments, an index is
determined that comprises a measure of myocardial perfusion and a
measure of MMP level based on the obtained images. In some
embodiments, the index derives from a logisitic regression analysis
of the summed rest score and the MMP level. In some embodiments,
the measure of myocardial perfusion is the summed stress score. In
some embodiments, the measure of perfusion is the summed rest
score. In some embodiments, the measure of perfusion is the summed
difference score.
[0020] In some embodiments, the MMP image and/or the myocardial
perfusion image is obtained before the subject exhibits any signs
of myocardial fibrosis. The presence of myocardial fibrosis may be
determined using, for example, delayed enhancement MRI of the heart
in whole or in part.
[0021] In another aspect, the invention provides a compound having
a structure selected from the group consisting of
##STR00004##
wherein F represents an isotopically-enriched population of
.sup.18F, and variants thereof comprising, instead of F, an
isotopically enriched imaging moiety selected from the group
consisting of .sup.11C, .sup.13N, .sup.123I, .sup.125I, .sup.99mTc,
.sup.95Tc, .sup.111In, .sup.62Cu, .sup.64Cu, .sup.67Ga, and
.sup.68Ga.
[0022] In another aspect, the invention provides a composition
comprising any of the foregoing compounds, and a pharmaceutically
acceptable carrier.
[0023] In another aspect, the invention provides use or methods of
use of the foregoing compounds in medical imaging techniques.
[0024] In another aspect, the invention provides methods for
determining the presence of an atherosclerotic plaque in a subject
comprising administering to a subject an imaging agent comprising
an MMP inhibitor linked to an imaging moiety and acquiring an image
of a portion of the subject (e.g., an image of the
cardiovasculature); and determining the presence of an
atherosclerotic plaque at least in part based on the presence of an
increased amount of imaging agent in the imaged portion of the
subject (for example, as compared to a region that does not contain
an atherosclerotic plaque).
[0025] In another aspect, the invention provides methods for
determining the effectiveness of an anti-lipid therapy on
atherosclerosis (e.g., on atherosclerotic plaque biology or
morphology) in a subject comprising administering to a subject a
first dose of an imaging agent comprising an MMP inhibitor linked
to an imaging moiety and acquiring at least one first image of a
portion of the subject; administering an anti-lipid therapy to the
subject; administering a second dose of an imaging agent comprising
an MMP inhibitor linked to an imaging moiety to the subject and
acquiring at least one second image of a portion of the subject;
determining the change in the amount of imaging agent in the
portion of the subject between the at least one first image and the
at least one second image; and determining the effectiveness of the
anti-lipid therapy based at least in part on the change in the
amount of the imaging agent in the portion of the subject between
the first image and the second image. The portion of the subject
being imaged is typically the cardiovasculature such as the
coronary arteries and the like.
[0026] In another aspect, the invention provides methods of
determining the presence of calcific aortic valve disease (CAVD) in
a subject and/or determining a subject's risk of developing CAVD
comprising administering to a subject an imaging agent comprising
an MMP inhibitor linked to an imaging moiety and acquiring a first
cardiac image of the subject; and determining the presence of CAVD
and/or a subject's risk of developing CAVD based at least in part
on the first cardiac image. The cardiac image typically includes or
is of the cardiac valve, optionally including the leaflets. In
accordance with the invention, increased uptake of the imaging
agent in the aortic valve region is indicative of the presence of
CAVD or the increased risk of developing CAVD. In some embodiments,
the subject being imaged does not have atherosclerosis (e.g., the
subject does not manifest symptoms associated with atherosclerosis,
and is referred to as "asymptomatic" for this condition). The
uptake of an MMP inhibitor linked to an imaging moiety in a
particular portion of a subject can allow for the imaging of tissue
remodeling characterized in part by macrophage infiltration, MMP
activation, and the like, in a portion of the subject, as a
diagnostic indicator of CAVD.
[0027] In another aspect, the invention provides methods of
determining the progression of CAVD in a subject comprising
administering to the subject a first dose of an imaging agent
comprising an MMP inhibitor linked to an imaging moiety and
acquiring at least one first cardiac image of the subject;
administering a second dose of an imaging agent comprising an MMP
inhibitor linked to an imaging moiety and acquiring at least one
second cardiac image of the subject; determining the change in the
amount of imaging agent in the heart or a portion of the heart
between the first image and the second image; and determining the
progression of CAVD in the subject based at least in part on the
change in the amount of the imaging agent in the heart or the
portion of the heart between the at least one first image and the
at least one second image. In important embodiments, the image
includes or is of the aortic valve, optionally including the
leaflets. In some embodiments, the method comprises administering
more than two doses of the imaging agent, obtaining more than two
cardiac images, and determining the progression of CAVD based at
least in part on the change in the amount of the imaging agent in
the heart or the portion of the heart between the obtained
images.
[0028] In another aspect, the invention provides methods of
determining the effectiveness of a treatment for CAVD in a subject
comprising administering to a subject a first dose of an imaging
agent comprising an MMP inhibitor linked to an imaging moiety and
acquiring at least one first cardiac image of the subject;
administering a treatment for CAVD to the subject; administering a
second dose of an imaging agent comprising an MMP inhibitor linked
to an imaging moiety and acquiring at least one second cardiac
image of the subject; determining the change in the amount of
imaging agent in the heart or a portion of the heart between the
first image and the second image; and determining the effectiveness
of the treatment for CAVD based at least in part on the change in
the amount of the imaging agent in the heart or a portion of the
heart between the at least one first image and the at least one
second image. In important embodiments, the image includes or is of
the aortic valve, optionally including the leaflets. The invention
contemplates that effectiveness of the treatment will be evidenced
by a decreased amount (and in some instances a constant or
unchanged amount) of imaging agent in the heart, and in particular
in the aortic valve region, optionally including the leaflets.
[0029] In various imaging methods set forth herein, two or more
images may be obtained from a subject. This may be the case where
for example the subject is undergoing a treatment and/or other
intervention between images or where it is desired to monitor the
progression of or towards a condition in the absence of treatment
or significant intervention. Treatment may be an anti-lipid
therapy, although it is not so limited. Intervention may be a
change in lifestyle, including for example weight loss, cessation
of smoking, increased exercise, and the like, although it too is
not so limited. When two or more images are obtained from a
subject, the images may be randomly spaced or regularly spaced in
time, including for example about weekly, biweekly, monthly,
bimonthly, every 6 months, or yearly.
[0030] These and other aspects of the invention will be described
in greater detail herein.
BRIEF DESCRIPTION OF DRAWINGS
[0031] FIG. 1. Comparison of myocardial .sup.261Tl and
.sup.99mTc-RP805 activity with MMP zymography in pig hearts 1, 2,
and 4 weeks post-MI. Shown are color coded spatial maps of relative
myocardial .sup.201Tl activity (top row), .sup.99mTc-RP805 activity
(second row), MMP-9 activity assessed by zymography (third row),
total and active MMP-2 activity assessed by zymography (fourth and
fifth rows), MMP-7 levels (sixth row), MT1-MMP levels (seventh
row), and MMP activity determined as a function of cleavage of a
fluorescent substrate (seventh row). Images are oriented with the
lateral wall on the right. Note the time dependent changes in
regional .sup.99mTc-RP805 activity correlate with spatial and
temporal changes in MMP levels/activity as assessed by zymography.
Reference color bar is at the bottom. Relationship between in vivo
and in vitro determination of MMP activity (upper right). In vivo
MMP activity, determined as retention of .sup.99mTc-RP805, was
significantly related to in vitro changes in levels of active MMP-2
(y=1687e.sup.0.033x, r: 0.89, p<0.05). (lower right) There was a
significant relationship between the change in body mass indexed LV
end-diastolic volume relative to control values and MMP activity
within the MI region (y=31.34e0.48x, r=0.38, p=0.04).
[0032] FIG. 2. In vivo (top) and ex vivo (middle) SPECT/CT images
were obtained in control pigs (n=7) and in pigs 10 days (n=2).
Increased MMP activation is seen in infarct region and in both
atria. Graph of myocardial RP805 uptake (% injected dose/g) for
control pigs and pigs 10 days and 4 weeks post-MI (bottom). There
were significant increases in .sup.99mTc-RP805 uptake within both
atria and infarct region at 10 days post-MI. At 4 weeks, post-MI
infarct region remained significantly elevated (defined as
p<0.003 after Bonferonni correction to increase stringency) as
did the LA region (p<0.01).
[0033] FIG. 3. Representative transaxial slices from ex vivo SPECT
images of a control pig heart, and hearts from pigs at 1 and 2
weeks post-MI. The top row of each image contain are targeted
.sup.99mTc-RP805 images (linear grey scale) matched with
corresponding high resolution CT images (gray scale, below). Hearts
were filled with alginate mixed with CT contrast to define right
and left ventricles (RV & LV) and atria (RA & LA). Uniform
uptake is seen in the control heart. Infarcted hearts demonstrate
focal .sup.99mTc-RP805 in the infarct region and in the atria.
[0034] FIG. 4. Results from quantitative analysis of ex vivo
.sup.99mTc-RP805 SPECT images from a pig heart at 4 weeks post-MI.
A. Uncorrected SPECT images, B. SPECT images with resolution
recovery, C. SPECT data reconstructed with partial volume
correction (PVC), D. Grey-scale .sup.99mTc-RP805 activity for 8
radial sectors per slice from gamma well counter, E. Postmortem
images of heart demonstrating dense inferolateral scar and marked
wall thinning, F. Correlation between measured regional myocardial
well-counter activity and SPECT derived activity with PVC.
[0035] FIG. 5. Plot of the MMP total activity per unit time,
according to some embodiments.
[0036] FIG. 6. Plot of the percent area collagen for the left and
right regions of the heart. Regional changes in matrix structure
could be detected by 1 week post-MI which were significant by 4
weeks post-MI (Kruskal-Wallis, p<0.05). In the inset: PSR
Imaging revealed matrix disruption and discontinuity between atrial
myocytes which was progressive with time post-MI.
[0037] FIG. 7. .sup.99mTc-RP805 in vivo microSPECT/CT imaging
(left) of MMP activation in an ApoE-/- mouse fed a Western diet for
9 months. Tracer uptake in the aortic valve area is indicated by
the arrows. Uptake in the aortic valve was confirmed by ex vivo
planar imaging (right) of the explanted heart and aorta.
[0038] FIG. 8. Plot of in vivo uptake over time of 99mTc-RP805 in
the aortic valve over time in ApoE-/- mice fed a Western diet at
various timepoints, and related data.
[0039] FIG. 9. Plot of ex vivo uptake of .sup.99mTc-RP805 in the
aortic valve over time in ApoE-/- mice fed a Western diet.
[0040] FIG. 10. Autoradiography of the explanted aorta from an
ApoE-/- mouse fed a Western diet for three months. Arrows indicate
uptake of .sup.111In-RP782 in the aortic valve area.
[0041] FIG. 11. H&E staining of the aortic valve in ApoE-/-
mice fed a Western diet for 4 (left) and 9 months (right)
demonstrating marked remodeling of valve leaflets over time.
[0042] FIG. 12. Immunostaining of F4-80 (dark grey) in the aortic
valve from an ApoE-/- mouse fed a Western diet for 6 months
demonstrating considerable macrophage infiltration.
[0043] FIG. 13. Plots of aortic valve GAPDH-normalized CD68 (top)
and MMP-12 (bottom) mRNA expression quantified by real time RT-PCR
in wild type (WT) mice on normal chow and ApoE-/- mice fed a
Western diet for 3, 6 or 9 months.
[0044] FIG. 14. Grey scale-coded non contrast CT images of an
ApoE-/- mouse fed a Western diet for 10 months demonstrating
calcification of the aortic valve. Arrows indicate the aortic valve
plane.
[0045] FIG. 15. M mode echocardiographic images of ApoE-/- mice fed
a Western diet demonstrating normal systolic separation of aortic
valve cusps after 3 months on diet (left) and reduced separation
after 9 months (right).
DETAILED DESCRIPTION OF INVENTION
[0046] The invention is based, in part, on the unexpected finding
that MMP levels, and in some instances more specifically
cardiovascular MMP levels, aortic valve MMP levels, and/or cardiac
MMP levels, can be observed using, for example, MMP inhibitors
linked to imaging moieties, and that such MMP levels can be used to
evaluate the presence of (including early detection of) or the
likelihood of developing certain conditions, the progression of the
condition, and the effectiveness of treatments and/or interventions
directed towards such conditions including prophylactic or
therapeutic treatments. The invention contemplates that increased
MMP levels (and thus increased MMP activity) is indicative of
tissue (e.g., vascular) remodeling observed in certain
cardiovascular conditions and/or that predisposes certain
cardiovascular conditions such as but not limited to AF.
[0047] Some aspects of the invention relate to determining the
presence of or the risk of developing AF. In some embodiments,
cardiac MMP levels are indicative of whether a subject is likely to
develop AF, either as a primary event or as a recurrence. The
methods of the invention can be used to identify subjects having an
increased risk (as compared to the risk of control or "normal"
subjects) of developing AF. Subjects identified in this manner may
then be monitored more closely (including more regularly) or they
may be treated at an earlier time point than previously
contemplated in order to reduce the risk that AF will ultimately
develop. In some embodiments, any increased MMP level relative to
control is used to identify subjects at increased risk. In some
embodiments, the MMP level (over control level) is used to quantify
the risk of developing AF. In these latter instances, lower MMP
levels may correlate with lower risk and higher MMP levels may
correlate with higher risk of AF. It is to be understood that MMP
levels may be indicated by retention levels of MMP imaging agents
of the invention.
[0048] The invention also contemplates a method for determining a
treatment regimen for a subject having AF as well as a method of
determining the likelihood of response to a treatment in a subject
having a history of AF. The method comprises administering to a
subject (having AF and/or having a history of AF) an imaging agent
of the invention and obtaining a heart (cardiac) image of the
subject, wherein the MMP level in the subject indicates whether the
subject should be treated using electrical cardioversion or an
alternative treatment such as pharmacological rate or rhythm
management, ablation and/or an implantable rate device. The
invention thus provides, in some embodiments, methods to direct the
clinical management strategy of such AF subjects between rate
control (including pharmacological rate control) or rhythm control
(including device-based and/or pharmacological rhythm control)
treatments.
[0049] The invention further contemplates a method for determining
the extent and complexity of ablation therapy that may be needed to
treat AF in a subject.
MMPs and AF
[0050] Atrial fibrosis is a hallmark of structural remodeling that
contributes to the AF substrate. Left atrial tissue has been shown
in patients and in animal models of AF to contain deposits of
fibrillar collagen and expansion of the extracellular matrix (ECM).
Histologically determined extent of fibrosis and ECM expansion has
been shown to correlate with AF persistence (Circ 2004;
109:363-368). Delayed enhancement MRI has been used to image left
atrial fibrosis and has shown more fibrosis in patients with
persistent AF compared to paroxysmal AF (Circ Cardiovasc Imaging
2010; 3:231-239). As significant fibrosis is relatively permanent,
there would be greater clinical benefit to detecting the presence
and extent of more proximate pathologies. The role that MMPs play
in ECM remodeling suggests their utility in this context. Indeed,
in a canine model of heart failure, administration of an MMP
inhibitor attenuated the vulnerability to AF and reduced atrial
fibrosis compared to control animals (J Cardiac Failure 2008;
14:768-776). In patients in whom pharmacologic or electrical
cardioversion was attempted, refractory AF was significantly
associated with elevated MMP-2 levels (Europace 2009; 11:332-337).
US 2011/0009861 discloses methods of predicting AF recurrence
comprising detecting specified amounts of MMPs in body fluids.
[0051] The methods of this invention provide non-invasive methods
to evaluate levels of MMPs in the atria and in so doing ensure the
relevant origin of the MMP. As MMPs play a role in numerous normal
and pathologic processes, MMP levels measured in a body fluid
cannot be assured to represent MMP levels in the atrium.
Additionally, the spatial extent of MMP activity, as may be
determined using the methods of this invention, will predict the
spatial extent of fibrosis. Such spatial extent cannot be
determined by a body fluid measure.
[0052] Provided are profiles of quantities, intensities,
concentrations, spatial distribution and/or localization of an
imaging agent comprising an MMP inhibitor that are an indication of
increased risk of developing primary or recurrent AF. The profiles
that are indication of higher risk of recurrence of AF in a subject
can be relative to a normal value. A normal value of an imaging
agent comprising an MMP inhibitor can be a reference value for an
age matched subject that is confirmed to have no evidence of
significant cardiovascular disease, or of AF. Thus, the normal
value can be a population-based value derived from a significant
number of healthy individuals. These reference normal values can be
obtained from population based studies.
[0053] Methods are also provided for determining whether a subject
with a history of AF should be treated with an implantable pacer.
These methods can comprise measuring retention of an imaging agent
comprising an MMP inhibitor in the subject to produce a retention
profile (that acts as a surrogate for MMP levels). The produced
profile can be used to determine, inter alia, whether the subject
will have a recurrence of AF following a therapy such as a
cardioversion procedure, a likely recurrence of AF indicating
treatment with an implantable pacer.
Prognosis
[0054] Provided herein is a method of predicting recurrence of AF
in a subject, comprising measuring retention of an imaging agent,
comprising an MMP inhibitor in a subject and comparing said levels
to reference values.
[0055] As used herein "recurrence" can include paroxysmal,
persistent, and chronic episodes of AF in a subject that has
experienced a prior episode of AF and may or may not have been
treated (including successfully treated in the short term) for the
prior episode of AF. Predicting recurrence of AF in a subject with
a history of (including presenting with) AF can be done prior to
administration of AF treatment and to determine if the selected AF
treatment will likely be followed by recurrence of AF in that
subject. For example, the recurrence of AF can be predicted prior
to electrical cardioversion. If it is determined that electrical
cardioversion will correct the AF without recurrence of AF then
electrical cardioversion can be selected as the treatment modality
of choice. If it is determined that electrical cardioversion will
not result in sustained AF correction, then another treatment
modality can be selected. For example, an implantable pacer device
can be selected or pharmacological pacing can be used instead of
electrical cardioversion if it is determined that there will be
recurrence with electrical cardioversion. Moreover, retention level
of an imaging agent comprising an MMP inhibitor can be used to
select treatment with an implantable pacer device, ablation, or by
pharmacological pacing.
[0056] The method of the invention can further comprise the steps
of measuring a ventricular perfusion defect and measuring uptake of
an imaging agent comprising an MMP inhibitor. Thus, the method can
comprise performing a resting flurpiridaz F 18 perfusion study,
determining the summed rest score from said study, quantifying
atrial uptake of an imaging agent comprising an MMP inhibitor and
determining risk of recurrent AF.
[0057] The invention further contemplates using cardiovascular MMP
levels to evaluate conditions other than AF. In some embodiments,
MMP levels may be useful for determining the presence of
atherosclerotic plaques in a subject and/or the effectiveness of
anti-lipid therapies (e.g., employed for the treatment of
atherosclerosis) on plaque biology in a subject. In some
embodiments, cardiac MMP levels (including levels in the aortic
valve region) can be used to determine the presence of CAVD in a
subject, to determine a subject's risk of developing CAVD, to
determine progression of CAVD in a subject, and/or to determine the
effectiveness of a treatment for CAVD in a subject. These and other
aspects and embodiments of the invention will be described in
greater detail herein.
MMPs and Atherosclerosis
[0058] Atherosclerosis, a major cause of morbidity and mortality in
the US, is linked to hyperlipidemia. Pharmacologic treatment of
hyperlipidemia is a common treatment for atherosclerotic diseases
and is believed to be related at least in part to "stabilizing"
effects on atherosclerotic plaque biology. The term
"atherosclerosis" is given its ordinary meaning in the art and
refers to a disease of the arterial wall in which the wall area
thickens, causing narrowing of the channel and thus impairing blood
flow. Atherosclerosis may occur in any area of the body, but can be
most damaging to a subject when it occurs in the heart, brain, or
blood vessels leading to the brain stem. Atherosclerosis includes
thickening and hardening of arterial walls or the accumulation of
fat, cholesterol and other substances that form atheromas or
plaques. Atherosclerosis may also result from calcification,
hemorrhage, ulceration, thrombosis, and/or trauma.
[0059] As noted above, in some embodiments, the invention provides
non-invasive methods to evaluate levels of MMPs in a portion of a
subject. In some embodiments, imaging of MMPs (e.g., MMP activation
in vivo) may be used to determine the presence of an
atherosclerotic plaque in a subject. In some embodiments, serial
imaging of MMPs (e.g., MMP activation in vivo) may be used to
determine the effectiveness of an anti-lipid therapy on plaque
biology in a subject. For example, a series of images may be
obtained during and/or following the course of administration of a
treatment for atherosclerosis to a subject, and the images may be
analyzed to determine effectiveness of the treatment. An
efficacious treatment is indicated by a reduced level of MMP
activation in the imaged region. An efficacious treatment may also
be indicated by an unchanged level of MMP activation in the imaged
region, in some instances. A non-efficacious treatment is indicated
by an increased level of MMP activation in the imaged region, in
some instances.
[0060] In some embodiments, a method of determining the presence of
an atherosclerotic plaque in a subject comprises administering to
the subject an imaging agent comprising an MMP inhibitor linked to
an imaging moiety and acquiring at least one first image of a
portion of the subject. The presence of an atherosclerostic plaque
may be determined based on the amount of imaging agent present in
the imaged portion of the subject. In some cases, the portion of
the subject is the heart or a portion of the heart (e.g., the
aortic arch).
[0061] In some embodiments, the location and/or concentration of
MMPs determined from the images may be analyzed relative to a
normal value. A normal value of an imaging agent of the invention
can be a reference value for an age-matched subject that is
confirmed to have no evidence of significant atherosclerosis. Thus,
the normal value can be a population-based value derived from a
significant number of healthy individuals. These reference normal
values can be obtained from population-based studies. The normal
value may be determined based on imaging agent uptake in the aortic
arch.
[0062] In some embodiments, a method of determining the
effectiveness of an anti-lipid therapy on plaque biology in a
subject comprises administering to the subject a first dose of an
imaging agent comprising an MMP inhibitor linked to an imaging
moiety and acquiring at least one first image of a portion of the
subject. An anti-lipid therapy may then be administered to the
subject. Following and/or concurrent to administration of the
anti-lipid therapy, the subject may be administered a second dose
of an imaging agent comprising an MMP inhibitor linked to an
imaging moiety and at least one second image of a portion of the
subject may be acquired. The change (e.g., decrease and/or
increase) in the amount of imaging agent in the portion of the
subject between the first image and the second image may be
determined. The effectiveness of the anti-lipid therapy may be
determined based at least in part on the change in the amount of
the imaging agent in the portion of the subject between the first
image and the second image. In some cases, a decrease in the amount
of imaging agent in the portion of the subject indicates that the
anti-lipid therapy is effective in reducing the amount of plaque in
the subject. In some cases, the portion of the subject is the heart
or a portion of the heart. In some cases, the change in the amount
of imaging agent in the aortic arch may be analyzed/determined. In
some cases, a lack of an increase in the amount of imaging agent in
the imaged portion of the subject (i.e., a constant level or a
decreased level) may also indicate an effective anti-lipid
therapy.
[0063] In some embodiments, the subject may be administered
additional doses of an imaging agent comprising an MMP inhibitor
linked to an imaging moiety and additional images of a portion of
the subject may be acquired. The change in the amount of imaging
agent in the portion of the subject between two images, or more
than two images, may be used to determine the effectiveness of the
anti-lipid therapy.
[0064] Those of ordinary skill in the art will be aware of
anti-lipid therapies which may be used to treat atherosclerosis in
a subject, for example, statins (e.g., rosuvastatin), fibrates
(e.g., gemfibrozil, fenofibrate), dietary changes, etc.
[0065] In some important embodiments, the imaging agent comprises
the structure:
##STR00005##
wherein F represents an isotopically-enriched population of
.sup.18F, and variants thereof comprising, instead of F, an
isotopically enriched imaging moiety selected from the group
consisting of .sup.11C, .sup.13N, .sup.123I, .sup.125I, .sup.99mTc,
.sup.95Tc, .sup.111In, .sup.62Cu, .sup.64Cu, .sup.67Ga, and
.sup.68Ga.
MMPs and Calcific Aortic Valve Disease (CAVD)
[0066] Calcific aortic valve disease (CAVD) is common among the
elderly population. Inflammation and matrix remodeling play a
central role in progression of CAVD to symptomatic aortic stenosis.
MMPs are upregulated in CAVD. Generally, the term "calcific aortic
valve disease" encompasses a disease spectrum from initial
alterations in the cell biology of the leaflets to end-stage
calcification resulting in left ventricular outflow obstruction.
Disease progression is generally characterized by a process of
thickening of the valve leaflets and the formation of calcium
nodules--often including the formation of actual bone--and new
blood vessels, which are concentrated near the aortic surface.
End-stage disease, e.g., calcific aortic stenosis, is generally
characterized pathologically by large nodular calcific masses
within the aortic cusps that protrude along the aortic surface into
the sinuses of Valsalva, interfering with opening of the cusps.
[0067] As noted above, in some embodiments, the invention provides
non-invasive methods to evaluate levels of MMPs in a portion of the
subject. In some embodiments, imaging of MMPs (e.g., MMP activation
in vivo) can be used to determine the presence of CAVD in a subject
(including early detection and/or diagnosis prior to irreversible
injury), to determine a subject's risk of developing CAVD, to
determine progression of CAVD in a subject, and/or to determine the
effectiveness of a treatment for CAVD in subject. For example, a
single image may be obtained of a subject, wherein the image may be
analyzed to determine the presence of CAVD in a subject and/or the
subject's risk of developing CAVD. As another example, a series of
images may be obtained of a subject over a period of time, wherein
the images may be analyzed to determining the progression of CAVD
in the subject and/or the effectiveness of a treatment for
CAVD.
[0068] In some embodiments, a method of determining the presence of
CAVD in a subject and/or determining a subject's risk of developing
CAVD comprises administering to the subject an imaging agent
comprising an MMP inhibitor linked to an imaging moiety and
acquiring a first cardiac image of the subject. The presence of
CAVD in the subject and/or the subject's risk of developing CAVD
may be based at least in part on the first cardiac image. In some
cases, the amount of imaging agent in the aortic valve may be
analyzed/determined. In some embodiments, the subject is
asymptomatic for atherosclerosis.
[0069] The images and related MMP values may be analyzed relative
to a normal value. A normal value of an imaging agent comprising an
MMP inhibitor can be a reference value for an age-matched subject
that is confirmed to have no evidence of significant CAVD. Thus,
the normal value can be a population-based value derived from a
significant number of healthy individuals. These reference normal
values can be obtained from population-based studies. The normal
value may be determined based on the level of imaging agent uptake
in a portion of the heart (e.g., aortic valve).
[0070] In some embodiments, the images obtained of a portion of a
subject (e.g., of a subject's aortic valve) may be analyzed in
connection with and/or with reference to other images or data
obtained from the subject. For example, in some cases, the images
obtained of a subject's aortic valve indicating MMP levels may be
analyzed in connection with and/or with reference to a CT scan of
the subject's aortic valve, wherein the CT may indicate the
presence or absence of calcification of the subject's aortic valve.
Other non-limiting examples including a subject's cholesterol
levels (e.g. LDL levels), blood pressure, and/or cardiac
dysfunction (e.g., as determined by ultrasound or EKG). In some
cases, the subject may have been diagnosed as having or as being at
risk of developing CAVD and/or atherosclerosis. In other cases, the
subject may be asymptomatic and/or may have not been diagnosed as
having CAVD and/or atherosclerosis. In some instances, the images
obtained using for example another imaging agent and/or another
modality may provide no evidence of calcification and thus the
subject may be one that manifests no calcification and yet is
experiencing the early stages of CAVD (i.e., precalcification
stages of CAVD).
[0071] In some embodiments, a method of determining the progression
of CAVD in a subject comprises administering to the subject a first
dose of an imaging agent comprising an MMP inhibitor linked to an
imaging moiety and acquiring at least one first cardiac image of
the subject. At a later time point, the subject may be administered
a second dose of an imaging agent comprising an MMP inhibitor
linked to an imaging moiety and at least one second cardiac image
of the subject may be acquired. The change (e.g., decrease and/or
increase) in the amount of imaging agent in the heart or a portion
of the heart between the first image and the second image may be
determined. The progression of CAVD in a subject may be determined
based at least in part on the change in the amount of the imaging
agent in the heart or a portion of the heart between the first
image and the second image. In some cases, the change in the amount
of imaging agent in the aortic valve may be
analyzed/determined.
[0072] In some embodiments, a method of determining the
effectiveness of a treatment for CAVD in a subject comprises
administering to a subject a first dose of an imaging agent
comprising an MMP inhibitor linked to an imaging moiety and
acquiring at least one first cardiac image of the subject. The
treatment for CAVD may then be administered to the subject.
Following and/or concurrent to administration of the treatment for
CAVD, the subject may be administered a second dose of an imaging
agent comprising an MMP inhibitor linked to an imaging moiety and
at least one second cardiac image of the subject may be acquired.
The change (e.g., decrease and/or increase) in the amount of
imaging agent in the heart or a portion of the heart between the
first image and the second image may be determined. The
effectiveness of the treatment for CAVD may be determined based at
least in part on the change in the amount of the imaging agent in
the heart or a portion of the heart between the first image and the
second image. In some cases, the lack of an increase in the amount
of the imaging agent in the imaged portions of the subject may
indicate effectiveness of the treatment. In some cases, the change
in the amount of imaging agent in the aortic valve may be
analyzed/determined.
[0073] In some embodiments, the imaging agent is .sup.99mTc-RP805
or .sup.111In-RP782. In some important embodiments, the imaging
agent comprises the structure:
##STR00006##
wherein F represents an isotopically-enriched population of
.sup.18F, and variants thereof comprising, instead of F, an
isotopically-enriched imaging moiety selected from the group
consisting of .sup.11C, .sup.13N, .sup.123I, .sup.125I, .sup.99mTc,
.sup.95Tc, .sup.111In, .sup.62Cu, .sup.64Cu, .sup.67Ga, and
.sup.68Ga.
Imaging Agents
[0074] The imaging agents of the invention comprise an MMP
inhibitor linked to an imaging moiety. Localization of the MMP
inhibitor (e.g., through binding to MMP resident in a tissue) is
indicative of the MMP level which in turn is indicative of MMP
activity. As will be understood, MMP inhibitor presence, location
and/or amount is detected by virtue of the imaging moiety linked to
the MMP inhibitor.
[0075] MMP inhibitors refer to agents that bind to one or more
MMPs. The MMPs may be but are not limited to MMP-2, MMP-9 and/or
MMP-14. Preferably, the MMP inhibitors bind to one or more MMPs for
a period of time that is sufficient to detect their presence in the
tissue being imaged.
[0076] As used herein, an "imaging moiety" refers to an atom or
group of atoms that is capable of producing a detectable signal
itself or upon exposure to an external source of energy (e.g.,
imaging agents comprising imaging moieties may allow for the
detection, imaging, and/or monitoring of the presence and/or
progression of a condition), pathological disorder, and/or disease.
Nuclear medicine imaging agents can include .sup.11C, .sup.13N,
.sup.18F, .sup.123I, .sup.125I, .sup.99mTc, .sup.95Tc, .sup.111In,
.sup.62Cu, .sup.64Cu, .sup.67Ga, and .sup.68Ga as the imaging
moiety. In some embodiments, the imaging moiety is .sup.18F.
[0077] In some embodiments, a compound (e.g., an imaging agent, a
fluoride species) may be isotopically-enriched with fluorine-18.
"Isotopically-enriched" refers to a composition containing isotopes
of an element such that the resultant isotopic composition is other
than the natural isotopic composition of that element. With regard
to the compounds provided herein, when a particular atomic position
is designated as .sup.19F, it is to be understood that the
abundance of .sup.19F at that position is substantially greater
than the natural abundance of .sup.18F, which is essentially zero.
In some embodiments, a fluorine designated as .sup.18F may have a
minimum isotopic enrichment factor of about 0.01%, about 0.05%,
about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about
0.75%, about 1%, about 2%, about 3%, about 4%, about 5%, about 10%,
about 15%, about 20%, about 30%, about 40%, about 50%, about 60%,
about 70%, about 80%, about 90%, about 95%, or greater. The
isotopic enrichment of the compounds provided herein can be
determined using conventional analytical methods known to one of
ordinary skill in the art, including mass spectrometry and
HPLC.
[0078] Imaging moieties include, without limitation, radioisotopes,
paramagnetic metal atoms, gas-filled microspheres, and the like. It
will be understood that the nature of the imaging moiety will
depend in large part on the imaging modality to be used. As an
example, imaging moieties that are radioisotopes are known to be
useful for imaging by gamma scintigraphy or positron emission
tomography (PET), and thus would be suitable if such imaging
modalities were being used in the methods of the invention. As
another example, the imaging moiety may be a metallic radioisotope
or a paramagnetic metal atom suitable for magnetic resonance
imaging (MRI). In these instances, the MMP inhibitor may be linked
to one or more chelators, and the imaging moiety binds to the
chelator(s). In these instances, the imaging moiety is
non-covalently linked to the MMP inhibitor via the chelator. As yet
another example, the MMP inhibitor may be linked to gas-filled
microspheres when ultrasound is the imaging modality of choice. The
MMP inhibitor may be linked to the material that encapsulates
and/or stabilizes such microspheres. In an important embodiment,
the imaging moiety is .sup.18F.
[0079] Each MMP inhibitor may be linked to one or more imaging
moieties. The MMP inhibitors may be directly or indirectly linked
to an imaging moiety. Indirect linkage may involve the use of a
linker or a spacer. Linkage may be covalent or non-covalent. In
important embodiments, covalent linkage is preferred.
[0080] Imaging agents suitable for use in the methods of the
invention are shown below along with their precursors:
TABLE-US-00001 Imaging Agent Precursors Imaging Agents ##STR00007##
##STR00008## ##STR00009## ##STR00010## ##STR00011## ##STR00012##
##STR00013## ##STR00014## ##STR00015## ##STR00016## ##STR00017##
##STR00018##
[0081] The imaging agents of Table 1 may be synthesized from the
precursors also shown in Table 1. LG refers to the leaving group
and may be but is not limited to tosylates and mesylates. Methods
for synthesizing imaging agents from tosylate or other precursor
forms are described in published PCT application WO
2011/097649.
[0082] Other imaging agents suitable for use in the methods of the
invention include those described in U.S. Pat. No. 6,656,448 and in
U.S. Pat. No. 6,989,139, the specific teachings of which are
incorporated by reference herein.
[0083] Those of ordinary skill in the art will be aware of methods
for synthesizing the imaging agents and imaging agent precursors
described herein. For example, see the methods disclosed in the
Examples section as well as those described in U.S. Pat. No.
6,656,448 and in U.S. Pat. No. 6,989,139, the specific teachings of
which are incorporated by reference herein.
[0084] Some imaging agents suitable for use in the methods of the
invention include those having one of the two following
structures:
##STR00019##
[0085] wherein, R is independently OH or --CH.sub.2SH; R.sup.1 is
independently selected at each occurrence from the group: H, OH,
C.sub.1-C.sub.3 alkyl, C.sub.2-C.sub.3 alkenyl, C.sub.2-C.sub.3
alkynyl, and heterocycle-S--CH.sub.2--; R.sup.2 is independently
C.sub.1-C.sub.20 alkyl; X is independently C.dbd.O or SO.sub.2
provided when X is C.dbd.O, R.sup.3 is
##STR00020##
[0086] and when X is SO.sub.2, R.sup.3 is independently selected
from the group: aryl substituted with 0-2 R.sup.6, and heterocycle
substituted with 0-2 R.sup.6; R.sup.4 is independently selected at
each occurrence from the group: C1-C6 alkyl, phenyl, and benzyl;
R.sup.5 is independently at each occurrence from the group:
[0087] NH(C1-C6 alkyl), NH-phenyl, and NH-heterocycle; wherein said
alkyl, phenyl and heterocycle groups are optionally substituted
with a bond to the linking group or a bond to the chelator; R.sup.6
is independently aryloxy substituted with 0-3 R.sup.7; R.sup.7 is
independently halogen or methoxy;
[0088] or alternatively, R.sup.1 and R.sup.4 may be taken together
to form a bridging group of the formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally substituted
with a bond to the linking group or a bond to the chelator; or
alternatively, R.sup.1 and R.sup.2 may be taken together to form a
bridging group of the formula --(CH.sub.2).sub.3--NH--, optionally
substituted with a bond to the linking group or a bond to the
chelator; or R.sup.1 and R.sup.2 taken together with the nitrogen
and carbon atom through which they are attached form a C.sub.5-7
atom saturated ring system substituted with one or more
substituents selected from the group consisting of: a bond to
L.sub.n, where L.sub.n is a linking group between the matrix
metalloproteinase inhibitor and chelator a bond to C.sub.h, and
--C.dbd.O--NR.sup.29R.sup.30; where C.sub.h is a chelator, R.sup.8
is independently at each occurrence OH or phenyl, optionally
substituted with a bond to the linking group or a bond to the
chelator, provided that when R.sup.8 is phenyl, R.sup.10 is
--C(.dbd.O)--CR.sup.12--NH--CH(CH.sub.3)--COOH; R.sup.9 and
R.sup.9' are independently H, C1-C6 alkyl optionally substituted
with a bond to the linking group or a bond to the chelator, or are
taken together with the carbon atom to which R.sup.9 and R.sup.9'
are attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-3 heteroatoms selected from O, N,
SO.sub.2 and S, said ring system substituted with R.sup.6 and
optionally substituted with a bond to the linking group or a bond
to the chelator; R.sup.10 and R.sup.11 are independently H, or
C1-C6 alkyl optionally substituted with a bond to the linking group
or a bond to the chelator, or are taken together with the nitrogen
atom to which they are attached to form a 5-7 atom saturated,
partially unsaturated or aromatic ring system containing 0-3
heteroatoms selected from O, N, SO.sub.2 and S, said ring system
optionally substituted with 0-3 R.sup.27, a bond to the linking
group or a bond to the chelator; or alternatively, R.sup.9 and
R.sup.10 are taken together with the carbon atom to which they are
attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-3 heteroatoms selected from O, N,
SO.sub.2 and S, said ring system optionally substituted with a bond
to the linking group or a bond to the chelator; and R.sup.12 is
independently C1-C20 alkyl; R.sup.27 is =0, C1-4 alkyl, or phenyl
substituted with R.sup.28; R.sup.28 is a phenoxy group substituted
with 0-2 OCH.sub.3 groups;
[0089] R.sup.29 and R.sup.30 taken together with the nitrogen atom
through which they are attached form a C5-7 atom saturated ring
system substituted with R.sup.31 and R.sup.31 is a benzyloxy group
substituted with C1-4 alkyl. In all of the foregoing embodiments, a
metallic imaging moiety is chelated by the chelator.
[0090] Still other imaging agents comprise (a) an imaging moiety
that is a diagnostic metal, and (b) a compound selected from [0091]
2-{[5-(3-{2-[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicyclo[10.-
2.2]hexadeca-1(15),12(16),13-triene-10-carbonyl)-amino]-acetylamino}propyl-
carbamoyl)-pyridin-2-yl]-hydrazonomethyl}-benzenesulfonic acid;
[0092]
2-{[5-(4-{[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicyclo[10.2.-
2]hexadeca-1(15),12(16),13-triene-10-carbonyl)-amino]-methyl}-benzylcarbam-
oyl)-pyridin-2-yl]-hydrazonomethyl}-benzene sulfonic acid; [0093]
2-[7-({N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl-
)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonyl--
amino}acetyl
amino)propyl]carbamoyl}methyl)-1,4,7,10-tetraaza-4,10-bis(carboxymethyl)c-
yclododecyl]acetic acid; [0094]
2-{7-[(N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-carbony
1-amino}methyl)phenyl]methyl}carbamoyl)methyl]-1,4,7,10-tetraaza-4,10-bis-
(carboxymethyl)cyclododecyl}acetic acid; [0095]
2-(7-{[N-(1-{N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-methylpropy-
l)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonyl-
-amino}acetylamino)propyl]carbamoyl}-2-sulfoethyl)carbamoyl]methyl}-1,4,7,-
10-tetraaza-4,10-bis(carboxymethyl)cyclododecyl)acetic acid; [0096]
2-[7-({N-[1-(N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylp-
ropyl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-y
1]carbonylamino}methyl)phenyl]methyl}carbamoyl)-2-sulfoethyl]carbamoyl}me-
thyl)-1,4,7,10-tetraaza-4,10-bis(carboxymethyl)cyclododecyl]acetic
acid; [0097]
2-({2-[({N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-met-
hyl propyl)-11-oxa-5-oxo
bicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylamino}acetylam-
ino)propyl]carbamoyl}methyl)(carboxymethyl)amino}ethyl){2-[bis(carboxymeth-
yl)amino]ethyl}amino]acetic acid; [0098]
2-[(2-{[(N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropy-
l)-11-oxa-5-oxo
bicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylamino}methyl)p-
henyl]methyl}carbamoyl)methyl](carboxymethyl)amino}ethyl){2-[bis(carboxyme-
thyl)amino]ethyl}amino]acetic acid; [0099]
N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-11-ox-
a-5-oxo
bicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonyl-amino}-
acetylamino)propyl]-4,5-bis[2-(ethoxyethyl-thio)acetylamino]pentanamide;
[0100]
N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxo
bicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylamino}methyl)--
phenyl]methyl}-4,5-bis[2-(ethoxyethylthio)acetylamino]-pentanamide;
[0101]
1-(1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamino).alpha.,.omega.-dicar-
bonylPEG3400-2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylam-
ino}-N-(3-aminopropyl)acetamide;
[0102]
1-(1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamino).alpha.,.omega.-
-dicarbonylPEG
3400-[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-11-oxa-5--
oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-N-{[4-(aminomethyl)-
phenyl]methyl}carboxamide conjugate; [0103]
2-[2-({5-[N-(5-(N-hydroxycarbamoyl)(5R)-5-{3-[4-(3,4-dimethoxyphenoxy)phe-
nyl]-3
methyl-2-oxopyrrolidinyl}pentyl)carbamoyl](2-pyridyl)}amino)(1Z)-2--
azavinyl]benzenesulfonic acid; and [0104]
2-(2-{[5-(N-{3-[3-(N-hydroxycarbamoyl)(4S)-4-({4-[(4-methylphenyl)methoxy-
]piperidyl}carbonyl)piperidyl]-3-oxopropyl}carbamoyl)(2-pyridyl)]amino}(1Z-
)-2-azavinyl)benzenesulfonic acid.
[0105] Still other examples of suitable imaging agents comprise an
imaging moiety such as a diagnostic metal attached to a compound
comprising either of the following structures:
##STR00021##
[0106] In still yet other embodiments, the imaging agent comprises
the structure:
##STR00022##
wherein F represents an isotopically-enriched population of
.sup.18F, and variants thereof comprising, instead of F, an
isotopically-enriched imaging moiety selected from the group
consisting of .sup.11C, .sup.13N, .sup.123I, .sup.125I, .sup.99mTc,
.sup.95Tc, .sup.111In, .sup.62Cu, .sup.64Cu, .sup.67Ga, and
.sup.68Ga. That is, wherein F is replaced with an
isotopically-enriched imaging moiety or a chelator associated with
an isotopically-enriched imaging moiety, wherein the imaging moiety
is selected from the group consisting of .sup.11C, .sup.13N,
.sup.123I, .sup.125I, .sup.99mTc, .sup.95Tc, .sup.111In, .sup.62Cu,
.sup.64Cu, .sup.67Ga, and .sup.68Ga. a. First Non-Limiting Set of
Embodiments of Imaging Agents or Precursors Thereof
[0107] This section provides non-limiting embodiments of compounds
which may function as imaging agents and/or imaging agent
precursors. In some embodiments, a compound is provided, wherein
the compound may be associated with a radioisotope (e.g., a
cytotoxic radioisotope), thereby forming an imaging agent.
(1) In some embodiments, the compound is of embodiment 1 of this
first non-limiting set of embodiments, wherein the compound
comprises: a) 1-10 targeting moieties b) a chelator (Ch); and c)
0-1 linking groups (Ln) between the targeting moiety and chelator;
wherein the targeting moiety is a matrix metalloproteinase
inhibitor; and wherein the chelator is capable of conjugating to a
cytotoxic radioisotope. (2) A compound according to embodiment 1,
wherein the targeting moiety is a matrix metalloproteinase
inhibitor having an Inhibitory constant K.sub.i of <1000 nM. (3)
A compound according to embodiment 1, wherein the targeting moiety
is a matrix, metalloproteinase inhibitor having an inhibitory
constant K.sub.i of <100 nM. (4) A compound according to any one
of embodiments 1-3, comprising 1-5 targeting moieties. (5) A
compound according to embodiment 1, comprising one targeting
moiety. (6) A compound according to any one of embodiments 1-5,
wherein the targeting moiety is a matrix metalloproteinase
inhibitor of the formulae (Ia) or (Ib):
##STR00023##
wherein, [0108] R is independently OH or --CH.sub.2SH; [0109]
R.sup.1 is independently selected at each occurrence from the
group: H, OH, C.sub.1-3 alkyl, C.sub.2-3 alkenyl, C.sub.2-3
alkynyl, and heterocycle-S--CH.sub.2--; [0110] R.sup.2 is
independently C.sub.1-20 alkyl; [0111] X is independently C.dbd.O
or SO.sub.2, provided when X is C.dbd.O, R.sup.3 is
##STR00024##
[0111] and when X is SO.sub.2, R.sup.3 is independently selected
from the group: aryl substituted with 0-2 R.sup.6, and heterocycle
substituted with 0-2 R.sup.6; [0112] R.sup.4 is independently
selected at each occurrence from the group: C.sub.1-6 alkyl,
phenyl, and benzyl; [0113] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NR-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to the linking group or a
bond to the chelator; [0114] R.sup.6 is independently aryloxy
substituted with 0-3 R.sup.7; [0115] R.sup.7 is independently
halogen or methoxy; [0116] or alternatively, [0117] R.sup.1 and
R.sup.4 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally
substituted with a bond to the linking group or a bond to the
chelator; [0118] or alternatively, [0119] R.sup.1 and R.sup.2 may
be taken together to form a bridging group of the formula
--(CH.sub.2).sub.3--NH--, optionally substituted with a bond to the
linking group or a bond to the chelator; or [0120] R.sup.1 and
R.sup.2 taken together with the nitrogen and carbon atom through
which they are attached form a C.sub.5-7 atom saturated ring system
substituted with one or more substituents selected from the group
consisting of: a bond to Ln, a bond to Ch, and
--C(.dbd.O)--NR.sup.29R.sup.30; [0121] R.sup.8 is independently at
each occurrence OH or phenyl, optionally substituted with a bond to
the linking group or a bond to the chelator, provided that when
R.sup.8 is phenyl, R.sup.10 is
--C(.dbd.O)--CR.sup.12--NH--CH(CH.sub.3)--COOH; [0122] R.sup.9 and
R.sup.9' are independently H, C.sub.1-6 alkyl optionally
substituted with a bond to the linking group or a bond to the
chelator, or are taken together with the carbon atom to which
R.sup.9 and R.sup.9' are attached to form a 5-7 atom saturated,
partially unsaturated or aromatic ring system containing 0-3
heteroatoms selected from O, N, SO.sub.2 and S, said ring system
substituted with R.sup.6 and optionally substituted with a bond to
the linking group or a bond to the chelator; [0123] R.sup.10 and
R.sup.11 are independently H, or C.sub.1-6 alkyl optionally
substituted with a bond to the linking group or a bond to the
chelator, or are taken together with the nitrogen atom to which
they are attached to form a 5-7 atom saturated, partially
unsaturated or aromatic ring system containing 0-3 heteroatoms
selected from O, N, SO.sub.2 and S, said ring system optionally
substituted with 0-3 R.sup.27, a bond to the linking group or a
bond to the chelator; [0124] or alternatively, [0125] R.sup.9 and
R.sup.10 are taken together with the carbon atom to which they are
attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-3 heteroatoms selected from O, N,
SO.sub.2 and S, said ring system optionally substituted with a bond
to the linking group or a bond to the chelator; and [0126] R.sup.12
is independently C.sub.1-20 alkyl; [0127] R.sup.27 is .dbd.O, C1-4
alkyl, or phenyl substituted with R.sup.28; [0128] R.sup.28 is a
phenoxy group substituted with 0-2 OCH.sub.3 groups; [0129]
R.sup.29 and R.sup.30 taken together with the nitrogen atom through
which they are attached form a C5-7 atom saturated ring system
substituted with R.sup.31: and [0130] R.sup.31 is a benzyloxy group
substituted with C1-4 alkyl. (7) A compound according to any one of
embodiments 1-6 wherein the targeting moiety is a matrix
metalloproteinase inhibitor of the formulae (Ia) or (Ib):
[0130] ##STR00025## [0131] R is OH; [0132] R.sup.1 is independently
selected at each occurrence from the group: H, OH, C.sub.1-3 alkyl,
C.sub.2-3 alkenyl, C.sub.2-3 alkynyl, and
heterocycle-S--CH.sub.2--; [0133] R.sup.2 is independently
C.sub.1-6 alkyl; [0134] X is C.dbd.O; [0135] R.sup.4 is
independently selected at each occurrence from the group: C.sub.1-6
alkyl, phenyl, and benzyl; [0136] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to the linking group or a
bond to the chelator; [0137] R.sup.6 is independently aryloxy
substituted with 0-3 R.sup.7; [0138] R.sup.7 is independently
halogen or methoxy; [0139] or alternatively, [0140] R.sup.1 and
R.sup.4 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally
substituted with a bond to the linking group or a bond to the
chelator; [0141] or alternatively, [0142] R.sup.1 and R.sup.2 may
be taken together to form a bridging group of the formula
--(CH.sub.2).sub.3--NH--, optionally substituted with a bond to the
linking group or a bond to the chelator; or [0143] R.sup.1 and
R.sup.2 taken together with the nitrogen and carbon atom through
which they are attached form a C.sub.5-7 atom saturated ring system
substituted with one or more substituents selected from the group
consisting of: a bond to Ln, a bond to Ch, and
--C(.dbd.O)--NR.sup.29R.sup.30; [0144] R.sup.8 is OH; [0145]
R.sup.9 and R.sup.9' are independently H, C.sub.1-6 alkyl
optionally substituted with a bond to the linking group or a bond
to the chelator, or are taken together with the carbon atom to
which R.sup.9 and R.sup.9' are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-1 heteroatoms selected from O, N, said ring system optionally
substituted with a bond to the linking group or a bond to the
chelator; [0146] R.sup.10 and R.sup.11 are independently H, or
C.sub.1-6 alkyl optionally substituted with a bond to the linking
group or a bond to the chelator, or are taken together with the
nitrogen atom to which they are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-1 heteroatoms selected from O, N, said ring system optionally
substituted with 0-3 R.sup.27, a bond to the linking group or a
bond to the chelator; [0147] or alternatively, [0148] R.sup.9 and
R.sup.10 are taken together with the carbon atom to which they are
attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-1 heteroatoms selected from O, N,
said ring system optionally substituted with a bond to the linking
group or a bond to the chelator; and [0149] R.sup.12 is
independently C.sub.1-6 alkyl; [0150] R.sup.27 is .dbd.O, C1-4
alkyl, or phenyl substituted with R.sup.28; [0151] R.sup.28 is a
phenoxy group substituted with 0-2 OCH.sub.3 groups; [0152]
R.sup.29 and R.sup.30 taken together with the nitrogen atom through
which they are attached form a C5-7 atom saturated ring system
substituted with R.sup.31; and [0153] R.sup.31 is a benzyloxy group
substituted with C1-4 alkyl. (8) A compound according to any one of
embodiments 1-7 wherein: [0154] R is --OH; [0155] R.sup.2 is
C.sub.1-6 alkyl; [0156] X is C.dbd.O; [0157] R.sup.3 is
[0157] ##STR00026## [0158] R.sup.1 and R.sup.4 are taken together
to form a bridging group of formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.2--; [0159] R.sup.5 is
NH(C1-6alkyl), substituted with a bond to the linking group or a
bond to the chelator. A compound according to any one of
embodiments 1-8, wherein: [0160] R is --OH; [0161] R.sup.9 is
C.sub.1 alkyl substituted with a bond to Ln; [0162] R.sup.10 and
R.sup.11 taken together with the nitrogen atom to which they are
attached form a 5 atom saturated ring system, said right system is
substituted with 0-3 R.sup.27; [0163] R.sup.27 is .dbd.O, C1-4
alkyl, or phenyl substituted with R.sup.28; and [0164] R.sup.28 is
a phenoxy group substituted with 0-2 OCH.sub.3 groups. (9) A
compound according to any one of embodiments 1-8, wherein: [0165] R
is --OH; [0166] R.sup.1 and R.sup.2 taken together with the
nitrogen and carbon atom through which they are attached form a
C.sub.5-7 atom saturated ring system substituted with one or more
substituents selected from the group consisting of: a bond to Ln, a
bond to Ch, and --C(.dbd.O)--NR.sup.29R.sup.30; [0167] R.sup.29 and
R.sup.30 taken together with the nitrogen atom through which they
are attached form a C5-7 atom saturated ring system substituted
with R.sup.31; and [0168] R.sup.31 is a benzyloxy group substituted
with C1-4 alkyl. (10) A compound according to any one of
embodiments 1-9, wherein the linking group is of the formula:
[0168]
((W.sup.1).sub.h-(CR.sup.13R.sup.14).sub.g).sub.x--(Z).sub.k--((C-
R.sup.13aR.sup.14a).sub.g'-(W.sup.2).sub.h').sub.x'; [0169] W.sup.1
and W.sup.2 are independently selected at each occurrence from the
group: O, S, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NBC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2, SO.sub.2NH,
--(OCH.sub.2CH.sub.2).sub.76-84, (OCR.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0170] aa is
independently at each occurrence an amino acids [0171] Z is
selected from the group: aryl substituted with 0-3 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.16; [0172] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, PO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-3 R.sup.16, aryl substituted with 0-3 R.sup.16,
benzyl substituted with 0-3 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-3 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to the chelator; [0173] R.sup.16 is independently
selected at each occurrence from the group: a bond to the chelator,
COOR.sup.17, C(.dbd.O)NHR.sup.17, NHC(.dbd.O)R.sup.17, OH,
NHR.sup.17, SO.sub.3H, PO.sub.3H, --OPO.sub.3H.sub.2, --OSO.sub.3H,
aryl substituted with 0-3 R.sup.17, C.sub.1-5 alkyl substituted
with 0-1 R.sup.18, C.sub.1-5 alkoxy substituted with 0-1 R.sup.18,
and a 5-10 membered heterocyclic ring system containing 1-4
heteroatoms independently selected from N, S, and O and substituted
with 0-3 R.sup.17; [0174] R.sup.17 is independently selected at
each occurrence from the group: H, alkyl substituted with 0-1
R.sup.18, aryl substituted with 0-1 R.sup.18, a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-1 R.sup.18,
C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.18, polyalkylene
glycol substituted with 0-1 R.sup.18, carbohydrate substituted with
0-1 R.sup.18, cyclodextrin substituted with 0-1 R.sup.18, amino
acid substituted with 0-1 R.sup.18, polycarboxyalkyl substituted
with 0-1 R.sup.18, polyazaalkyl substituted with 0-1 R.sup.18,
peptide substituted with 0-1 R.sup.18, wherein the peptide is
comprised of 2-10 amino acids, 3,6-O-disulfo-B-D-galactopyranosyl,
bis(phosphonomethyl)glycine, and a bond to the chelator; [0175]
R.sup.18 is a bond to the chelator; [0176] k is selected from 0, 1,
and 2; [0177] h is selected from 0, 1, and 2; [0178] h' is selected
from 0, 1, and 2; [0179] g is selected from 0, 1, 2, 3, 4, 5, 6, 7,
8, 9, and 10; [0180] g' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8,
9, and 10; [0181] s is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,
and 10; [0182] s' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,
and 10; [0183] s'' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,
and 10; [0184] t is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and
10; [0185] t' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and
10; [0186] x is selected from 0, 1, 2, 3, 4, and 5; and [0187] x'
is selected from 0, 1, 2, 3, 4, and 5. 11. A compound according to
any one of embodiments 6-10 wherein [0188] W.sup.1 and W.sup.2 are
independently selected at each occurrence from the group: O, NH,
NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O), C(.dbd.O)NR.sup.15,
C(.dbd.O), C(.dbd.O)O, OC(.dbd.O), NBC(.dbd.S)NH, NHC(.dbd.O)NH,
SO.sub.2, --(CH.sub.2CH.sub.2O).sub.76-84--,
(OCH.sub.2CH.sub.2).sub.s, (CH.sub.2CH.sub.2O).sub.s',
(OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0189] aa is
independently at each occurrence an amino acid; [0190] Z is
selected from the group: aryl substituted with 0-1 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-1
R.sup.16; [0191] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-1 R.sup.16, aryl substituted with 0-1 R.sup.16,
benzyl substituted with 0-1 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-1 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to the chelator; [0192] k is 0 or 1; [0193] s is
selected from 0, 1, 2, 3, 4, and 5; [0194] s' is selected from 0,
1, 2, 3, 4, and 5; [0195] s'' is selected from 0, 1, 2, 3, 4, and
5; and [0196] t is selected from 0, 1, 2, 3, 4, and 5. (12) A
compound according to any one of embodiments 6-11, wherein: [0197]
W.sup.1 is C(.dbd.O)NR.sup.15; [0198] h is 1; [0199] g is 3; [0200]
R.sup.13 and R.sup.14 are independently H; [0201] x is 1; [0202] k
is 0; [0203] g' is 0; [0204] h' is 1; [0205] W.sup.2 is NH; and
[0206] x' is 1. (13) A compound according to any one of embodiments
6-12, wherein: [0207] x is 0; [0208] k is 1; [0209] Z is aryl
substituted with 0-3 R.sup.16; [0210] g' is 1; [0211] W.sup.2 is
NH; [0212] R.sup.13a and R.sup.14a are independently H; [0213] h'
is 1; and [0214] x' is 1. (14) A compound according to any one of
embodiments 6-13, wherein: [0215] W.sup.1 is C(.dbd.O)NR.sup.15;
[0216] h is 1; [0217] g is 2; [0218] R.sup.13 and R.sup.14 are
independently H; [0219] x is 1, [0220] k is 0; [0221] g' is 1;
[0222] R.sup.13a and R.sup.14a are independently H; or C1-5 alkyl
substituted with 0-3 R.sup.16; [0223] R.sup.16 is SO.sub.3H; [0224]
W.sup.2 is NHC(.dbd.O) or NH; [0225] h' is 1; and [0226] x' is 2.
(15) A compound according to any one of embodiments 6-14, wherein:
[0227] W.sup.1 is C(.dbd.O)NH; [0228] h is 1; [0229] g is 3; [0230]
R.sup.13 and R.sup.14 are independently H; [0231] k is 0; [0232] g'
is 0; [0233] x is 1; [0234] W.sup.2 is --NH(C.dbd.O)-- or
--(OCH.sub.2CH.sub.2).sub.76-84--; [0235] h' is 2; and [0236] x' is
1. (16) A compound according to any one of embodiments 6-15,
wherein: [0237] x is 0; [0238] k is 0; [0239] g' is 3; [0240] h' is
1; [0241] W.sup.2 is NH; and [0242] x' is 1. (17) A compound
according to any one of embodiments 6-16, wherein: [0243] x is 0;
[0244] z is aryl substituted with 0-3 R.sup.16; [0245] k is 1;
[0246] g' is 1; [0247] R.sup.13aR.sup.14a are independently H;
[0248] W.sup.2 is NHC(.dbd.O) or --(OCH2CH2).sub.76-84--; and
[0249] x' is 1. (18) A compound according to any one of embodiments
6-17, wherein: [0250] W.sup.1 is C.dbd.O; [0251] g is 2; [0252]
R.sup.13 and R.sup.14 are independently H; [0253] k is 0; [0254] g'
is 0; [0255] h' is 1; [0256] W.sup.2 is NH; and [0257] x' is 1.
(19) A compound according to embodiment 1 wherein the linking group
is absent. (20) A compound according to any one of embodiments
6-19, wherein the chelator is a metal bonding unit having a formula
selected from the group:
[0257] ##STR00027## [0258] A.sup.1, A.sup.2, A.sup.3, A.sup.4,
A.sup.5, A.sup.6, A.sup.7, and A.sup.8 are independently selected
at each occurrence from the group: N, NR.sup.26, NR.sup.19,
NR.sup.19R.sup.20, S, SH, --S(Pg), O, OH, PR.sup.19,
PR.sup.19R.sup.20, --O--P(O)(R.sup.21)--O--, P(O)R.sup.21R.sup.22,
a bond to the targeting moiety and a bond to the linking group;
[0259] Pg is a thiol protecting group; [0260] E.sup.1, E.sup.2,
E.sup.3, E.sup.4, E.sup.5, E.sup.6, E.sup.7, and E.sup.8 are
independently a bond, CH, or a spacer group independently selected
at each occurrence from the group: C.sub.1-C.sub.16 alkyl
substituted with 0-3 R.sup.23, aryl substituted with 0-3 R.sup.23,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.23,
heterocyclo-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, wherein
the heterocyclo group is a 5-10 membered heterocyclic ring system
containing 1-4 heteroatoms independently selected from N, S, and O,
C.sub.6-10 aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-10 alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, and
a 5-10 membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23; [0261] R.sup.19 and R.sup.20 are each independently
selected from the group: a bond to the linking group, a bond to the
targeting moiety, hydrogen, C.sub.1-C.sub.10 alkyl substituted with
0-3 R.sup.23, aryl substituted with 0-3 R.sup.23, C.sub.1-10
cycloalkyl substituted with 0-3 R.sup.23, heterocyclo-C.sub.1-10
alkyl substituted with 0-3 R.sup.23, wherein the heterocyclo group
is a 5-10 membered heterocyclic ring system containing 1-4
heteroatoms independently selected from N, S, and O, C.sub.6-10
aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, C.sub.1-10
alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23, and an electron, provided that when one of R.sup.19 or
R.sup.20 is an electron, then the other is also an electron; [0262]
R.sup.21 and R.sup.22 are each independently selected from the
group: a bond to the linking group, a bond to the targeting moiety,
--OH, C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23, aryl
substituted with 0-3 R.sup.23, C.sub.3-10 cycloalkyl substituted
with 0-3 R.sup.23, heterocyclo-C.sub.1-10 alkyl substituted with
0-3 R.sup.23, wherein the heterocyclo group is a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O, C.sub.6-10 aryl-C.sub.1-10 alkyl
substituted with 0-3 R.sup.23, C.sub.1-10 alkyl-C.sub.6-10
aryl-substituted with 0-3 R.sup.23, and a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-3 R.sup.23; [0263]
R.sup.23 is independently selected at each occurrence from the
group: a bond to the linking group, a bond to the targeting moiety,
.dbd.O, F, Cl, Br, I, --CF.sub.3, --CN, --CO.sub.2R.sup.24,
--C(.dbd.O)R.sup.24, --C(.dbd.O)N(R.sup.24).sub.2, --CHO,
--CH.sub.2OR.sup.24, --OC(.dbd.O)R.sup.24, --OC(.dbd.O)OR.sup.24a,
--OR.sup.24, --OC(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25C(O)R.sup.24, --NR.sup.25C(.dbd.O)OR.sup.24a,
--NR.sup.25C(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25SO.sub.2N(R.sup.24).sub.2, --NR.sup.25SO.sub.2R.sup.24a,
--SO.sub.3H, --SO.sub.2R.sup.24a, --SR.sup.24,
--S(.dbd.O)R.sup.24a, --SO.sub.2N(R.sup.24).sub.2,
--N(R.sup.24).sub.2, --NHC(.dbd.S)NHR.sup.24, --NOR.sup.24,
NO.sub.2, --C(.dbd.O)NHOR.sup.24, --C(.dbd.O)NHNR.sup.24R.sup.24a,
--OCH.sub.2CO.sub.2H, 2-(1-morpholino)ethoxy, C.sub.1-C.sub.5
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 cycloalkylmethyl, C.sub.2-C.sub.6 alkoxyalkyl, aryl
substituted with 0-2 R.sup.24, and a 5-10 membered heterocyclic
ring system containing 1-4 heteroatoms independently selected from
N, S, and O, and [0264] wherein at least one of A.sup.1, A.sup.2,
A.sup.3, A.sup.4, A.sup.5, A.sup.6, A.sup.7, A.sup.8 or R.sup.23 is
a bond to the linking group or targeting moiety; [0265] R.sup.24,
R.sup.24a, and R.sup.25 are independently selected at each
occurrence from the group: a bond to the linking group, a bond to
the targeting moiety, H, C.sub.1-C.sub.6 alkyl, phenyl, benzyl,
C.sub.1-C.sub.6 alkoxy, halide, nitro, cyano, and trifluoromethyl;
and [0266] R.sup.26 is a co-ordinate bond to a metal or a hydrazine
protecting group. (21) A compound according to any one of
embodiments 6-20 wherein: [0267] A.sup.1, A.sup.2, A.sup.3,
A.sup.4, A.sup.5, A.sup.6, A.sup.7, and A.sup.8 are independently
selected at each occurrence from the group: NR.sup.19,
NR.sup.19R.sup.20, S, SH, OH, a bond to the targeting moiety and a
bond to the linking group; [0268] E.sup.1, E.sup.2, E.sup.3,
E.sup.4, E.sup.5, E.sup.6, E.sup.7, and E.sup.8 are independently a
bond, CH, or a spacer group independently selected at each
occurrence from the group: C.sub.1-C.sub.10 alkyl substituted with
0-3 R.sup.23, aryl substituted with 0-3 R.sup.23, C.sub.3-10
cycloalkyl substituted with 0-3 R.sup.23, and a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-3 R.sup.23; [0269]
wherein at least one of A.sup.1, A.sup.2, A.sup.3, A.sup.4,
A.sup.5, A.sup.6, A.sup.7, A.sup.8 and R.sup.23 is a bond to the
linking group or a targeting moiety; [0270] R.sup.19, and R.sup.20
are each independently selected from the group: a bond to the
targeting moiety, a bond to the linking group, hydrogen,
C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23, aryl
substituted with 0-3 R.sup.23, a 5-10 membered heterocyclic ring
system containing 1-4 heteroatoms independently selected from N, S,
and O and substituted with 0-3 R.sup.23, and an electron, provided
that when one of R.sup.19 or R.sup.20 is an electron, then the
other is also an electron; [0271] R.sup.23 is independently
selected at each occurrence from the group: a bond to the targeting
moiety, a bond to the linking group, .dbd.O, F, C, Br, I,
--CF.sub.3, --CN, --CO.sub.2R.sup.24, --C(.dbd.O)R.sup.24,
--C(.dbd.O)N(R.sup.24).sub.2, --CH.sub.2OR.sup.24,
--OC(.dbd.O)R.sup.24, --OC(.dbd.O)OR.sup.24a, --OR.sup.24,
--OC(.dbd.O)N(R.sup.24).sub.2, --NR.sup.25C(.dbd.O)R.sup.24,
--NR.sup.25C(.dbd.O)OR.sup.24a,
--NR.sup.25C(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25SO.sub.2N(R.sup.24).sub.2, --NR.sup.25SO.sub.2R.sup.24a,
--SO.sub.3H, --SO.sub.2R.sup.24a, --S(.dbd.O)R.sup.24a,
--SO.sub.2N(R.sup.24).sub.2, --N(R.sup.24).sub.2,
--NHC(.dbd.S)NHR.sup.24, .dbd.NOR.sup.18,
--C(.dbd.O)NHNR.sup.18R.sup.18a, --OCH.sub.2CO.sub.2, and
2-(1-morpholino) ethoxy; and [0272] R.sup.24, R.sup.24a, and
R.sup.25 are independently selected at each occurrence from the
group: a bond to the linking group, H, and C.sub.1-C.sub.6 alkyl.
(22) A compound according to any one of embodiments 6-21 wherein
the chelator is of the formula:
[0272] ##STR00028## [0273] A.sup.1 is a bond to the linking group;
[0274] A.sup.2, A.sup.4, and A.sup.6 are each N; [0275] A.sup.3,
A.sup.5, A.sup.7 and A.sup.8 are each OH; [0276] E.sup.1, E.sup.2,
and E.sup.4 are C2 alkyl; [0277] E.sup.3, E.sup.5, E.sup.7, and
E.sup.8 are C.sub.2 alkyl substituted with 0-1 R.sup.23; [0278]
R.sup.23 is .dbd.O; (23) A compound according to any one of
embodiments 6-22 wherein the chelator is of the formula: [0279]
C.sub.h is
##STR00029##
[0279] wherein: [0280] A5 is a bond to Ln; [0281] A.sup.1, A.sup.3,
A.sup.7 and A.sup.8 are each OH; [0282] A.sup.2, A.sup.4 and
A.sup.6 are each NH; [0283] E.sup.1, E.sup.3, E.sup.5, E.sup.7, and
E.sup.8 are C.sub.2 alkyl substituted with 0-1 R.sup.23; [0284]
E.sup.2, and E.sup.4, are C.sub.2 alkyl; [0285] R.sup.23 is .dbd.O.
(24) A compound according to any one of embodiments 6-23 wherein
the chelator is of the formula:
[0285] ##STR00030## [0286] A.sup.1, A.sup.2, A.sup.3 and A.sup.4
are each N; [0287] A.sup.5, A.sup.6 and A.sup.8 are each OH; [0288]
A.sup.7 is a bond to L.sub.n; [0289] E.sup.1, E.sup.2, E.sup.3,
E.sup.4 are each independently C.sub.2 alkyl; and [0290] E.sup.5,
E.sup.6, E.sup.7, E.sup.8 are each independently C.sub.2 alkyl
substituted with 0-1 R.sup.23; [0291] R.sup.23 is .dbd.O. (25) A
compound according to any one of embodiments 6-24 wherein the
chelator is of the formula:
[0291] ##STR00031## [0292] A.sup.1 is NR.sup.26 [0293] R.sup.26 is
a co-ordinate bond to a metal or a hydrazine protecting group;
[0294] E.sup.1 is a bond; [0295] A.sup.2 is NHR.sup.19; [0296]
R.sup.19 is a heterocycle substituted with R.sup.23, the
heterocycle being selected from pyridine and pyrimidine; [0297]
R.sup.23 is selected from a bond to the linking group,
C(.dbd.O)NHR.sup.24 and C(.dbd.O)R.sup.24; and [0298] R.sup.24 is a
bond to the linking group. (26) A compound according to any one of
embodiments 6-25 wherein the chelator is of the formula:
##STR00032##
[0298] wherein: [0299] A.sup.1 and A.sup.3 are each --S(Pg); [0300]
Pg is a thiol protecting group [0301] E.sup.1 and E.sup.4 are
C.sub.2 alkyl substituted with 0-1 R.sup.23; [0302] R.sup.23 is
.dbd.O; [0303] A.sup.2 and A.sup.4 are each --NH; [0304] E.sup.2 is
CH.sub.2; [0305] E.sup.3 is C.sub.1-3 alkyl substituted with 0-1
R.sup.23; [0306] A.sup.3 is a bond to Ln. (27) A compound according
to any one of embodiments 6-26 wherein the chelator is of the
formula:
##STR00033##
[0306] wherein: [0307] A.sup.1 is a bond to Ln; [0308] E.sup.1 is
C.sub.1 alkyl substituted by R.sup.23; [0309] A.sup.2 is NH; [0310]
E.sup.2 is C.sub.2 alkyl substituted with 0-1R.sup.23; [0311]
A.sup.3 is --O--P(O)(R.sup.21)--O; [0312] E.sup.3 is C.sub.1 alkyl;
[0313] A.sup.4 and A.sup.5 are each --O--; [0314] E.sup.4 and
E.sup.6 are each independently C.sub.1-15 alkyl substituted with
0-1R.sup.23; [0315] E.sup.5 is C.sub.1 alkyl; [0316] R.sup.21 is
--OH; and [0317] R.sup.23 is .dbd.O. (28) A compound of embodiment
1 having the formula:
[0317] (Q).sub.d-L.sub.n-C.sub.h
wherein, Q is a compound of Formulae (Ia) or (Ib):
##STR00034##
wherein, [0318] R is independently OH or --CH.sub.2SH; [0319]
R.sup.1 is independently selected at each occurrence from the
group: H, OH, C.sub.1-3 alkyl, C.sub.2-3 alkenyl, C.sub.2-3
alkynyl, and heterocycle-S--CH.sub.2--; [0320] R.sup.2 is
independently C.sub.1-20 alkyl; [0321] X is independently C.dbd.O
or SO.sub.2, provided when X is C.dbd.O, R.sup.3 is
##STR00035##
[0321] and when X is SO.sub.2, R.sup.3 is independently selected
from the group: aryl substituted with 0-2 R.sup.6, and heterocycle
substituted with 0-2 R.sup.6; [0322] R.sup.4 is independently
selected at each occurrence from the group: C.sub.1-6 alkyl,
phenyl, and benzyl; [0323] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, pbenyl and heterocycle groups
are optionally substituted with a bond to L.sub.n; [0324] R.sup.6
is independently aryloxy substituted with 0-3 R.sup.7; [0325]
R.sup.7 is independently halogen or methoxy; [0326] or
alternatively, [0327] R.sup.1 and R.sup.4 may be taken together to
form a bridging group of the formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.3--, optionally substituted
with a bond to L.sub.n; [0328] or alternatively, [0329] R.sup.1 and
R.sup.2 may be taken together to form a bridging group of the
formula --(CH.sub.3).sub.3--NH--, optionally substituted with a
bond to L.sub.n; or [0330] R.sup.1 and R.sup.2 taken together with
the nitrogen and carbon atom through which they are attached form a
C.sub.5-7 atom saturated ring system substituted with one or more
substituents selected from the group consisting of: a bond to Ln, a
bond to Ch, and --C(.dbd.O)--NR.sup.29R.sup.30; [0331] R.sup.8 is
independently at each occurrence OH or phenyl, optionally
substituted with a bond to L.sub.n, provided that when R.sup.8 is
phenyl, R.sup.10 is --C(.dbd.O)--CR.sup.12--NH--CH(CH.sub.3)--COOH;
[0332] R.sup.9 and R.sup.9' are independently H, C.sub.1-6 alkyl
optionally substituted with a bond to L.sub.n, or are taken
together with the carbon atom to which they are attached to form a
5-7 atom saturated, partially unsaturated or aromatic ring system
containing 0-3 heteroatoms selected from O, N, SO.sub.2 and S, said
ring system substituted with R.sup.6 and optionally substituted
with a bond to L.sub.n; [0333] R.sup.10 and R.sup.11 are
independently H, or C.sub.1-6 alkyl optionally substituted with a
bond to La, or are taken together with the nitrogen atom to which
they are attached to form a 5-7 atom saturated, partially
unsaturated or aromatic ring system containing 0-3 heteroatoms
selected from O, N, SO.sub.2 and S, said ring system optionally
substituted with 0-3 R.sup.27 or a bond to L.sub.n; [0334] or
alternatively, [0335] R.sup.9 and R.sup.10 are taken together with
the carbon atom to which they are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-3 heteroatoms selected from O, N, SO.sub.2 and S, said ring
system optionally substituted with a bond to Ln; [0336] R.sup.12 is
independently C.sub.1-20 alkyl; [0337] d is selected from 1, 2, 3,
4, 5, 6, 7, 8, 9, and 10; [0338] L.sub.n is a linking group having
the formula:
[0338]
((W.sup.1).sub.h-(CR.sup.13R.sup.14).sub.g).sub.x--(Z).sub.k--((C-
R.sup.13aR.sup.14a).sub.g'-(W.sup.2).sub.h').sub.x'; [0339] W.sup.1
and W.sup.2 are independently selected at each occurrence from the
group: O, S, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2, SO.sub.2NH,
--(OCH.sub.2CH.sub.2).sub.76-84, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0340] aa is
independently at each occurrence an amino acid; [0341] Z is
selected from the group: aryl substituted with 0-3 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.16; [0342] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, PO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-3 R.sup.16, aryl substituted with 0-3 R.sup.16,
benzyl substituted with 0-3 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-3 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to C.sub.h; [0343] R.sup.16 is independently selected at
each occurrence from the group: a bond to C.sub.h, COOR.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)R.sup.17, OH, NHR.sup.17,
SO.sub.3H, PO.sub.3H, --OPO.sub.3H.sub.2, --OSO.sub.3H, aryl
substituted with 0-3 R.sup.17, C.sub.1-5 alkyl substituted with 0-1
R.sup.18, C.sub.1-5 alkoxy substituted with 0-1 R.sup.18, and a
5-10 membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.17; [0344] R.sup.17 is independently selected at each
occurrence from the group: H, alkyl substituted with 0-1 R.sup.18,
aryl substituted with 0-1 R.sup.18, a 5-10 membered heterocyclic
ring system containing 1-4 heteroatoms independently selected from
N, S, and O and substituted with 0-1 R.sup.18, C.sub.3-10
cycloalkyl substituted with 0-1 R.sup.18, polyalkylene glycol
substituted with 0-1 R.sup.18, carbohydrate substituted with 0-1
R.sup.18, cyclodextrin substituted with 0-1 R.sup.18, amino acid
substituted with 0-1 R.sup.18, polycarboxyalkyl substituted with
0-1 R.sup.18, polyazaalkyl substituted with 0-1 R.sup.18, peptide
substituted with 0-1 R.sup.18, wherein the peptide is comprised of
2-10 amino acids, 3,6-O-disulfo-B-D-galactopyranosyl,
bis(phosphonomethyl)glycine, and a bond to C.sub.h; [0345] R.sup.18
is a bond to C.sub.h; [0346] k is selected from 0, 1, and 2; [0347]
h is selected from 0, 1, and 2; [0348] h' is selected from 0, 1,
and 2, 3, 4, 5, 6, 7, 8, 9, and 10; [0349] g is selected from 0, 1,
2, 3, 4, 5, 6, 7, 8, 9, and 10; [0350] g' is selected from 0, 1, 2,
3, 4, 5, 6, 7, 8, 9, and 10; [0351] s is selected from 0, 1, 2, 3,
4, 5, 6, 7, 8, 9, and 10; [0352] s' is selected from 0, 1, 2, 3, 4,
5, 6, 7, 8, 9, and 10; [0353] s'' is selected from 0, 1, 2, 3, 4,
5, 6, 7, 8, 9, and 10; [0354] t is selected from 0, 1, 2, 3, 4, 5,
6, 7, 8, 9, and 10; [0355] t' is selected from 0, 1, 2, 3, 4, 5, 6,
7, 8, 9, and 05; [0356] x is selected from 0, 1, 2, 3, 4, and 5;
[0357] x' is selected from 0, 1, 2, 3, 4, and 5; [0358] C.sub.h is
a metal bonding unit having a formula selected from the group:
[0358] ##STR00036## [0359] A.sup.1, A.sup.2, A.sup.3, A.sup.4,
A.sup.5, A.sup.6, A.sup.7, and A.sup.8 are independently selected
at each occurrence from the group: N, NR.sup.26, NR.sup.19,
NR.sup.19R.sup.20, S, SH, --S(Pg), O, OH, PR.sup.19,
PR.sup.19R.sup.20, --O--P(O)(R.sup.21)--O--, P(O)R.sup.21R.sup.22,
a bond to the targeting moiety and a bond to the linking group;
[0360] Pg is a thiol protecting group; [0361] E.sup.1, E.sup.2,
E.sup.3, E.sup.4, E.sup.5, E.sup.6, E.sup.7, and E.sup.8 are
independently a bond, CH, or a spacer group independently selected
at each occurrence from the group: C.sub.1-C.sub.16 alkyl
substituted with 0-3 R.sup.23, aryl substituted with 0-3 R.sup.23,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.23,
heterocyclo-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, wherein
the heterocyclo group is a 5-10 membered heterocyclic ring system
containing 1-4 heteroatoms independently selected from N, S, and O,
C.sub.6-10 aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-10 alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, and
a 5-10 membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23; [0362] R.sup.19 and R.sup.20 are each independently
selected from the group: a bond to the linking group, a bond to the
targeting moiety, hydrogen, C.sub.1-C.sub.10 alkyl substituted with
0-3 R.sup.23, aryl substituted with 0-3 R.sup.23, C.sub.1-10
cycloalkyl substituted with 0-3 R.sup.23, heterocyclo-C.sub.1-10
alkyl substituted with 0-3 R.sup.23, wherein the heterocyclo group
is a 5-10 membered heterocyclic ring system containing 1-4
heteroatoms independently selected from N, S, and O, C.sub.6-10
aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, C.sub.1-10
alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23, and an electron, provided that when one of R.sup.19 or
R.sup.20 is an electron, then the other is also an electron; [0363]
R.sup.21 and R.sup.22 are each independently selected from the
group: a bond to the linking group, a bond to the targeting moiety,
--OH, C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23, aryl
substituted with 0-3 R.sup.23, C.sub.3-10 cycloalkyl substituted
with 0-3 R.sup.23, heterocyclo-C.sub.1-10 alkyl substituted with
0-3 R.sup.23, wherein the heterocyclo group is a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O, C.sub.6-10 aryl-C.sub.1-10 alkyl
substituted with 0-3 R.sup.23, C.sub.1-10 alkyl-C.sub.6-10
aryl-substituted with 0-3 R.sup.23, and a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-3 R.sup.23; [0364]
R.sup.23 is independently selected at each occurrence from the
group: a bond to the linking group, a bond to the targeting moiety,
.dbd.O, F, Cl, Br, I, --CF.sub.3, --CN, --CO.sub.2R.sup.24,
--C(.dbd.O)R.sup.24, --C(.dbd.O)N(R.sup.24).sub.2, --CHO,
--CH.sub.2OR.sup.24, --OC(.dbd.O)R.sup.24, --OC(.dbd.O)OR.sup.24a,
--OR.sup.24, --OC(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25C(.dbd.O)R.sup.24, --NR.sup.25C(.dbd.O)OR.sup.24a,
--NR.sup.25C(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25SO.sub.2N(R.sup.24).sub.2, --NR.sup.25SO.sub.2R.sup.24a,
--SO.sub.3H, --SO.sub.2R.sup.24a, --SR.sup.24,
--S(.dbd.O)R.sup.24a, --SO.sub.2N(R.sup.24).sub.2,
--N(R.sup.24).sub.2, --NHC(.dbd.S)NHR.sup.24, .dbd.NOR.sup.24,
NO.sub.2, --C(.dbd.O)NHOR.sup.24, --C(.dbd.O)NHNR.sup.24R.sup.24a,
--OCH.sub.2CO.sub.2H, 2-(1-morpholino)ethoxy, C.sub.1-C.sub.5
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 cycloalkylmethyl, C.sub.2-C.sub.6 alkoxyalkyl, aryl
substituted with 0-2 R.sup.24, and a 5-10 membered heterocyclic
ring system containing 1-4 heteroatoms independently selected from
N, S, and O; and [0365] wherein at least one of A.sup.1, A.sup.2,
A.sup.3, A.sup.4, A.sup.5, A.sup.6, A.sup.7, A.sup.8 or R.sup.23 is
a bond to the linking group or targeting moiety; [0366] R.sup.24,
R.sup.24a, and R.sup.25 are independently selected at each
occurrence from the group: a bond to the linking group, a bond to
the targeting moiety, H, C.sub.1-C.sub.6 alkyl, phenyl, benzyl,
C.sub.1-C.sub.6 alkoxy, halide, nitro, cyano, and trifluoromethyl;
and [0367] R.sup.26 is a co-ordinate bond to a metal or a hydrazine
protecting group; or [0368] a pharmaceutically acceptable salt
thereof. (29) A compound according to embodiment 28 wherein: [0369]
R is --OH; [0370] R.sup.2 is C.sub.1-6 alkyl; [0371] X is C.dbd.O:
[0372] R.sup.3 is
[0372] ##STR00037## [0373] R.sup.1 and R.sup.4 are taken together
to form a bridging group of formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.2--; [0374] R.sup.5 is
NH(C.sub.1-6alkyl), substituted with a bond to the linking group or
a bond to the chelator. (30) A compound according to any one of
embodiments 28-29 wherein [0375] R is --OH; [0376] R.sup.9 is
C.sub.1 alkyl substituted with a bond to Ln; [0377] R.sup.10 and
R.sup.11 taken together with the nitrogen atom to which they are
attached form a 5 atom saturated ring system, said right system is
substituted with 0-3 R.sup.27; [0378] R.sup.27 is .dbd.O, C1-4
alkyl, or phenyl substituted with R.sup.28; and [0379] R.sup.28 is
a phenoxy group substituted with 0-2 OCH.sub.3 groups. (31) A
compound according to any one of embodiments 28-30 wherein [0380] R
is --OH; [0381] R.sup.1 and R.sup.2 taken together with the
nitrogen and carbon atom through which they are attached form a
C.sub.5-7 atom saturated ring system substituted with one or more
substituents selected from the group consisting of: a bond to Ln, a
bond to Ch, and --C(.dbd.O)--NR.sup.29R.sup.30; [0382] R.sup.29 and
R.sup.30 taken together with the nitrogen atom through which they
are attached form a C5-7 atom saturated ring system substituted
with R.sup.31; and [0383] R.sup.31 is a benzyloxy group substituted
with C1-4 alkyl. (32) A compound according to any one of
embodiments 28-31 wherein [0384] d is selected from 1, 2, 3, 4, and
5; [0385] W is independently selected at each occurrence from the
group: O, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0386] aa is
independently at each occurrence an amino acid; [0387] Z is
selected from the group: aryl substituted with 0-1 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-1
R.sup.16; [0388] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-1 R.sup.16, aryl substituted with 0-1 R.sup.16,
benzyl substituted with 0-1 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-1 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to C.sub.h; [0389] k is 0 or 1; [0390] s is selected
from 0, 1, 2, 3, 4, and 5; [0391] s' is selected from 0, 1, 2, 3,
4, and 5; [0392] s'' is selected from 0, 1, 2, 3, 4, and 5; [0393]
t is selected from 0, 1, 2, 3, 4, and 5; [0394] A.sup.1, A.sup.2,
A.sup.3, A.sup.4, A.sup.5, A.sup.6, A.sup.7, and A.sup.8 are
independently selected at each occurrence from the group:
NR.sup.19, NR.sup.19R.sup.20, S, SH, OH, and a bond to L.sub.n;
[0395] E is a bond, CH, or a spacer group independently selected at
each occurrence from the group: C.sub.1-C.sub.10 alkyl substituted
with 0-3 R.sup.23, aryl substituted with 0-3 R.sup.23, C.sub.3-10
cycloalkyl substituted with 0-3 R.sup.23, and a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-3 R.sup.23; [0396]
R.sup.19, and R.sup.20 are each independently selected from the
group: a bond to Ln, hydrogen, C.sub.1-C.sub.10 alkyl substituted
with 0-3 R.sup.23, aryl substituted with 0-3 R.sup.23, a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23, and an electron, provided that when one of R.sup.19 or
R.sup.20 is an electron, then the other is also an electron; [0397]
R.sup.23 is independently selected at each occurrence from the
group; a bond to L.sub.n, .dbd.O, F, Cl, Br, I, --CF.sub.3, --CN,
--CO.sub.2R.sup.24, --C(.dbd.O)R.sup.24,
--C(.dbd.O)N(R.sup.24).sub.2, --CH.sub.2OR.sup.24,
--OC(.dbd.O)R.sup.24, --OC(.dbd.O)OR.sup.24a, --OR.sup.24,
--OC(.dbd.O)N(R.sup.24).sub.2, --NR.sup.25C(.dbd.O)R.sup.24,
--NR.sup.25C(.dbd.O)OR.sup.24a,
--NR.sup.25C(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25SO.sub.2N(R.sup.24).sub.2, --NR.sup.25SO.sub.2R.sup.24a,
--SO.sub.3H, --SO.sub.2R.sup.24a, --S(.dbd.O)R.sup.24a,
--SO.sub.2N(R.sup.24).sub.2, --N(R.sup.24).sub.2,
--NHC(.dbd.S)NHR.sup.24, .dbd.NOR.sup.18,
--C(.dbd.O)NHNR.sup.18R.sup.18a, --OCH.sub.2CO.sub.2H, and
2-(1-morpholino)ethoxy; and [0398] R.sup.24, R.sup.24a, and
R.sup.25 are independently selected at each occurrence from the
group: a bond to L.sub.n, H, and C.sub.1-C.sub.6 alkyl; and (33) A
compound according to any one of embodiments 28-32 wherein [0399] d
is 1, [0400] C.sub.h is
[0400] ##STR00038## [0401] A.sup.1 is a bond to L.sub.n; [0402]
A.sup.2, A.sup.4, and A.sup.6 are each N; [0403] A.sup.3, A.sup.5,
A.sup.7 and A.sup.8 are each OH; [0404] E.sup.1, E.sup.2, and
E.sup.4 are C2 alkyl; [0405] E.sup.3, E.sup.5, E.sup.7, and E.sup.8
are C.sub.2 alkyl substituted with 0-1 R.sup.23; [0406] R.sup.23 is
.dbd.O; (34) A compound according to any one of embodiments 28-33
wherein [0407] C.sub.h is
##STR00039##
[0407] wherein: [0408] A5 is a bond to Ln; [0409] A.sup.1, A.sup.3,
A.sup.7 and A.sup.8 are each OH; [0410] A.sup.2, A.sup.4 and
A.sup.6 are each NH; [0411] E.sup.1, E.sup.3, E.sup.5, E.sup.7, and
E.sup.8 are C.sub.2 alkyl substituted with 0-1 R.sup.23; [0412]
E.sup.2, and E.sup.4, are C.sub.2 alkyl; [0413] R.sup.23 is .dbd.O.
(35) A coepound according to any one of embodiments 28-34 is
wherein [0414] C.sub.h is
[0414] ##STR00040## [0415] A.sup.1, A.sup.2, A.sup.3 and A.sup.4
are each N; [0416] A.sup.5, A.sup.6 and A.sup.8 are each OH: [0417]
A.sup.7 is a bond to L.sub.n; [0418] E.sup.1, E.sup.2, E.sup.3,
E.sup.4 are each independently, C.sub.2 alkyl; and [0419] E.sup.5,
E.sup.6, E.sup.7, E.sup.8 are each independently, C.sub.2 alkyl
substituted with 0-1 R.sup.23; [0420] R.sup.23 is .dbd.O; (36) A
compound according to any one of embodiments 28-35 wherein [0421]
C.sub.h is
[0421] ##STR00041## [0422] A.sup.1 is NR.sup.26; [0423] R.sup.26 is
a co-ordinate bond to a metal; or a hydrazine protecting group;
[0424] E.sup.1 is a bond; [0425] A.sup.2 is NHR.sup.19; [0426]
R.sup.19 is a heterocycle substituted with R.sup.23, the
heterocycle being selected from pyridine and pyrimidine; [0427]
R.sup.23 is selected from a bond to L.sub.n, C(.dbd.O)NHR.sup.24
and C(.dbd.O)R.sup.24; and [0428] R.sup.24 is a bond to L.sub.n.
(37) A compound according to any one of embodiments 28-36
wherein
##STR00042##
[0428] wherein: [0429] A.sup.1 and A.sup.5 are each --S(Pg); [0430]
Pg is a thiol protecting group; [0431] E.sup.1 and E.sup.4 are
C.sub.2 alkyl substituted with 0-1 R.sup.23; [0432] R.sup.23 is
.dbd.O; [0433] A.sup.2 and A.sup.4 are each --NH; [0434] E.sup.2 is
CH.sub.2; [0435] E.sup.3 is C1-3 alkyl substituted with 0-1
R.sup.23; [0436] A.sup.3 is a bond to Ln. (38) A compound according
to any one of embodiments 28-37 wherein
##STR00043##
[0436] wherein [0437] A.sup.1 is a bond to Ln; [0438] E.sup.1 is
C.sub.1 alkyl substituted by R.sup.23; [0439] A.sup.2 is NH; [0440]
E.sup.2 is C.sub.2 alkyl substituted with 0-1R.sup.23; [0441]
A.sup.3 is --O--P(O)(R.sup.21)--O; [0442] E.sup.3 is C.sub.1 alkyl;
[0443] A.sup.4 and A.sup.5 are each --O--; [0444] E.sup.4 and
E.sup.6 are each independently C.sub.1-16 alkyl substituted with
0-1R.sup.23; [0445] E.sup.5 is C.sub.1 alkyl; [0446] A.sup.5 is
--O-- [0447] R.sup.21 is --OH; and [0448] R.sup.23 is .dbd.O. (39)
A compound according embodiment 28 wherein [0449] W.sup.1 is
C(.dbd.O)NR.sup.15; [0450] h is 1; [0451] g is 3; [0452] R.sup.13
and R.sup.14 are independently H; [0453] x is 1; [0454] k is 0:
[0455] g' is 0; [0456] h' is 1; [0457] W.sup.2 is NH; and [0458] x'
is 1. (40) A compound according to embodiments 28 wherein [0459] x
is 0; [0460] k is 1; [0461] Z is aryl substituted with 0-3
R.sup.16; [0462] g' is 1; [0463] W.sup.2 is NH; [0464] R.sup.13a
and R.sup.14a are independently H; [0465] h' is 1; and [0466] x' is
1. (41) A compound according to embodiments 28 wherein [0467]
W.sup.1 is C(O)NR.sup.15; [0468] h is 1; [0469] g is 2; [0470]
R.sup.13 and R.sup.14 are independently H; [0471] x is 1; [0472] k
is 0; [0473] g' is 1; [0474] R.sup.13a and R.sup.14a are
independently H; or C1-5 alkyl substituted [0475] with 0-3
R.sup.16; [0476] R.sup.16 is SO.sub.3H; [0477] W.sup.2 is
NHC(.dbd.O) or NH; [0478] h' is 1; and [0479] x' is 2. (42) A
compound according to embodiment 28 wherein [0480] W.sup.1 is
C(.dbd.O)NH; [0481] h is 1; [0482] g is 3; [0483] R.sup.13 and
R.sup.14 are independently H; [0484] k is 0; [0485] g' is 0; [0486]
x is 1; [0487] W.sup.2 is --NH(C.dbd.O)-- or
--(OCH.sub.2CH.sub.2).sub.76-84--; [0488] h' is 2; and [0489] x' is
1. (43) A compound according to embodiment 28 wherein [0490] x is
0; [0491] k is 0; [0492] g' is 3; [0493] h' is 1; [0494] W.sup.2 is
NH; and [0495] x' is 1. (44) A compound according to embodiment 28
wherein [0496] x is 0; [0497] Z is aryl substituted with 0-3
R.sup.16; [0498] k is 1; [0499] g' is 1; [0500] R.sup.13aR.sup.14a
are independently H; [0501] W.sup.2 is NHC(.dbd.O) or
--(OCH2CH2).sub.76-84--; and [0502] x' is 1. (45) A compound
according to embodiment 28 wherein [0503] W.sup.1 is C.dbd.O;
[0504] g is 2; [0505] R.sup.13 and R.sup.14 are independently H;
[0506] k is 0; [0507] g' is 0; [0508] h' is 1; [0509] W.sup.2 is
NH; and [0510] x' is 1. (46) A compound according to embodiment 1
or 28 selected from the group consisting of: [0511]
2-{[5-(3-{2-[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicy-
clo[10.2.2]hexadeca-1(15),12(16),13-triene-10-carbonyl)-amino]-acetylamino-
}-propylcarbamoyl)-pyridin-2-yl]-hydrazonomethyl}-benzenesulfonic
acid; [0512]
2-{[5-(4-{[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicycl-
o[10.2.2]hexadeca-1(15),12(16),13-trien-10-carbonyl)-amino]-methyl)-benzyl-
carbamoyl}-pyridin-2-yl]-hydrazonomethyl}-benzenesulfonic acid;
[0513]
2-[7-({N-[3-(2-{[7-(N-hydroxycabamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylam-
ino}acetylamino)propyl]carbamoyl}methyl)-1,4,7,10-tetraaza-4,10-bis(carbox-
ymethyl)cyclododecyl]acetic acid, [0514]
2-{7-[(N-{(4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-carbonyla-
mino{methyl)phenyl]methyl}carbamoyl)methyl]-1,4,7,10-tetraaza-4,10-bis(car-
boxymethyl)cyclododecyl}acetic acid; [0515]
2-(7-{[N-(1-{N-[3-(2-([7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methyl-
propyl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1
(15),12(16),13-trien-3-yl]carbonylamino)acetylamino)propyl]carbamoyl}-2-s-
ulfoethyl)carbamoyl]methyl}-1,4,7,10-tetraaza-4,10-bis(carboxymethyl)cyclo-
dodecyl)acetic acid; [0516]
2-[7-({N-[1-(N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylp-
ropyl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12
(16),13-trien-3-yl]-carbonylamino}methyl)phenyl]methyl}carbamoyl)-2-sulfo-
ethyl]carbamoyl}methyl)-1,4,7,10-tetraaza-4,10-bis(carboxymethyl)cyclodode-
cyl]acetic acid; [0517]
2-({2-[({N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylprop-
yl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbony-
lamino}acetylamino)propyl]carbamoyl}methyl)(carboxymethyl)amino}ethyl){2-[-
bis(carboxymethyl)amino]ethyl}amino]acetic acid; [0518]
2-[(2-{[(N-{4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl-
)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-carbonyl-
amino}methyl)phenyl]methyl}carbamoyl)methyl](carboxymethyl)amino}ethyl){2--
[bis(carboxymethyl)amino]ethyl}amino]acetic acid; [0519]
N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-11-ox-
a-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylamino}ac-
etylamino)propyl]-4,5-bis[2-(ethoxyethylthio)acetylamino]pentanamide;
[0520]
N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl)carbonylam-
ino}methyl)-phenyl]methyl}-4,5-bis[2-(ethoxyethylthio)acetylamino]-pentana-
mide; [0521]
1-(1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamino)-.alpha.,.omega.-dica-
rbonylPEG.sub.3400-2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylp-
ropyl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carb-
onylamino}-N-(3-aminopropyl)acetamide; [0522]
1-(1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamino)-.alpha.,.omega.-dica-
rbonylPEG.sub.3400-[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylprop-
yl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-N-{[4--
(aminomethyl)phenyl]methyl}carboxamide conjugate; [0523]
2-[2-({5-[N-(5-(N-hydroxycarbamoyl)(5R)-5-{3-[4-(3,4-dimethoxyphenoxy)phe-
nyl]-3-methyl-2-oxopyrrolidinyl}pentyl)carbamoyl](2-pyridyl)}amino)(1Z)-2--
azavinyl]benzenesulfonic acid; [0524]
2-(2-{[5-(N-{3-[3-(N-hydroxycarbamoyl)(4S)-4-({4-[(4-methylphenyl)methoxy-
]piperidyl}carbonyl)piperidyl]-3-oxopropyl}carbamoyl)(2-pyridyl)]amino}(1Z-
)-2-azavinyl)benzenesulfonic acid; and
##STR00044##
[0524] (47) In some embodiments, a radiopharmaceutical comprising a
compound of any one of embodiments 1-46 and a cytotoxic
radioisotope which is complexed to the chelator. (48) In some
embodiments, a radiopharmaceutical comprising a compound of any one
of embodiments 1-47 and a cytotoxic radioisotope which is complexed
to the chelator. (49) In some embodiments, a radiopharmaceutical
comprising a compound of any one of embodiments 1-47 and a
cytotoxic radioisotope. (50) In some embodiments, a
radiopharmaceutical according to embodiment 20 selected from the
group consisting of:
2-{[5-(3-{2-[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicyclo[10.-
-2.2]hexadeca-1(15),12(16),13-triene-10-carbonyl)-amino]-acetylamino}-prop-
y-lcarbamoyl)-pyridin-2-yl]-hydrazonomethyl}-benzenesulfonic acid;
and
2-{[5-(4-{[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicyclo[10.2.-
-2]hexadeca-1(15),12(16),13-triene-10-carbonyl)-amino]-methyl}-benzylcarba-
m-oyl)-pyridin-2-yl]-hydrazonomethyl}-benzenesulfonic acid; wherein
the cytotoxic radioisotope is .sup.99mTc. (51) In some embodiments,
a radiopharmaceutical according to embodiment 47 wherein the
cytotoxic radioisotope is selected from the group consisting of
beta particle emitters, alpha particle emitters, and Auger electron
emitters. (52) In some embodiments, a radiopharmaceutical according
to embodiment 47 wherein the cytotoxic radioisotope is selected
from the group consisting of: .sup.186Re, .sup.188Re, .sup.153Sm,
.sup.166Ho, .sup.177Lu, .sup.149Pm, .sup.90Y, .sup.212Bi,
.sup.103Pd, .sup.109Pd, .sup.159Gd, .sup.140La, .sup.198Au,
.sup.199Au, .sub.169Yb, .sup..sup.175Yb, .sup.165Dy, .sup.166Dy,
.sup.67Cu, .sup.105Rh, .sup.111Ag, and .sup.192Ir. (53) In some
embodiments, a radiopharmaceutical according to embodiment 47
wherein the cytotoxic radioisotope is selected from the group
consisting of: .sup.186Re, .sup.188Re, .sup.153Sm, .sup.166Ho,
.sup.177Lu, .sup.149Pm, .sup.90Y, .sup.212Bi, .sup.103Pd,
.sup.105Rh, (54) In some embodiments, a radiopharmaceutical
according to embodiment 47 wherein the cytotoxic radioisotope is
selected from the group consisting of: .sup.186Re, .sup.188Re,
.sup.153Sm, .sup.166Ho, .sup.177Lu, .sup.149Pm, .sup.90Y, and
.sup.212Bi. (55) In some embodiments, a composition comprising a
compound of any one of embodiments 1-54, or a pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable carrier.
b. Second Non-Limiting Set of Embodiments of Imaging Agents or
Precursors Thereof
[0525] This section provides non-limiting embodiments of compounds
which may function as imaging agents and/or imaging agent
precursors (also referred to herein as a diagnostic agent). In some
embodiments, a compound is provided, wherein the compounds may be
associated with a radioisotope, thereby forming an imaging agent
(or diagnostic agent).
(1) In some embodiments, the diagnostic agent (or imaging agent) is
of embodiment 1 of this second non-limiting set of embodiments,
wherein the diagnostic agent (or imaging agent) comprises: [0526]
i) 1-10 targeting moieties; [0527] ii) a chelator; and [0528] iii)
0-1 linking groups between the targeting moiety and chelator;
wherein the targeting moiety is a matrix metalloproteinase
inhibitor; and wherein the chelator is capable of conjugating to
the diagnostic metal. (2) A diagnostic agent according to
embodiment 1, wherein the targeting moiety is a matrix
metalloproteinase inhibitor having an inhibitory constant K.sub.i
of <1000 nM. (3) A diagnostic agent according to any of
embodiments 1-2, wherein the targeting moiety is a matrix
metalloproteinase inhibitor having an inhibitory constant K.sub.i
of <100 nM. (4) A diagnostic agent according to any of
embodiments 1-3, comprising 1-5 targeting moieties. (5). A
diagnostic agent according to any of embodiments 1-4, comprising
one targeting moiety. (6) A diagnostic agent any of embodiments
1-5, wherein the targeting moiety is an inhibitor of one or more
matrix metalloproteinases selected from the group consisting of
MMP-1, MMP-2, MMP-3, MMP-9, and MMP-14. (7) A diagnostic agent of
any one of embodiments 1-6, wherein the targeting moiety is an
inhibitor of one or more matrix metalloproteinases selected from
the group consisting of MMP-2, MMP-9, and MMP-14. (8) A diagnostic
agent according to any one of embodiments 1-7 wherein the targeting
moiety is a matrix metalloproteinase inhibitor of the formulae (Ia)
or (Ib):
##STR00045##
[0528] wherein, [0529] R is independently OH or --CH.sub.3SH;
[0530] R.sup.1 is independently selected at each occurrence from
the group: H, OH, C.sub.1-3 alkyl, C.sub.2-3 alkenyl, C.sub.2-3
alkynyl, and heterocycle-S--CH.sub.2--; [0531] R.sup.2 is
independently C.sub.1-20 alkyl; [0532] X is independently C.dbd.O
or SO.sub.2, provided when X is C.dbd.O, R.sup.3 is
##STR00046##
[0532] and when X is SO.sub.2, R.sup.3 is independently selected
from the group: aryl substituted with 0-2 R.sup.6, and heterocycle
substituted with 0-2 R.sup.6; [0533] R.sup.4 is independently
selected at each occurrence from the group: C.sub.1-6 alkyl,
phenyl, and benzyl; [0534] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to the linking group or a
bond to the chelator; [0535] R.sup.6 is independently aryloxy
substituted with 0-3 R.sup.7; [0536] R.sup.7 is independently
halogen or methoxy; [0537] or alternatively, [0538] R.sup.1 and
R.sup.4 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally
substituted with as bond to the linking group or a bond to the
chelator; [0539] or alternatively, [0540] R.sup.1 and R.sup.2 may
be taken together to form a bridging group of the formula
--(CH.sub.2).sub.3--NH--, optionally substituted with a bond to the
linking group or a bond to the chelator; or [0541] R.sup.1 and
R.sup.2 taken together with the nitrogen and carbon atom through
which they are attached form a C.sub.5-7 atom saturated ring system
substituted with one or more substituents selected from the group
consisting of: a bond to Ln, a bond to Ch, and
--C(.dbd.O)--NR.sup.29R.sup.30; [0542] R.sup.8 is independently at
each occurrence OH or phenyl, optionally substituted with a bond to
the linking group or a bond to the chelator, provided that when
R.sup.8 is phenyl, R.sup.10 is
--C(.dbd.O)--CR.sup.12--NH--CH(CH.sub.3)--COOH; [0543] R.sup.9 and
R.sup.9' are independently H, C.sub.1-6 alkyl optionally
substituted with a bond to the linking group or a bond to the
chelator, or are taken together with the carbon atom to which
R.sup.9 and R.sup.9' are attached to form a 5-7 atom saturated,
partially unsaturated or aromatic ring system containing 0-3
heteroatoms selected from O, N, SO.sub.2 and S, said ring system
substituted with R.sup.6 and optionally substituted with a bond to
the linking group or a bond to the chelator; [0544] R.sup.10 and
R.sup.11 are independently H, or C.sub.1-6 alkyl optionally
substituted with a bond to the linking group or a bond to the
chelator, or are taken together with the nitrogen atom to which
they are attached to form a 5-7 atom saturated, partially
unsaturated or aromatic ring system containing 0-3 heteroatoms
selected from O, N, SO.sub.2 and S, said ring system optionally
substituted with 0-3 R.sup.27, a bond to the linking group or a
bond to the chelator; [0545] or alternatively, [0546] R.sup.9 and
R.sup.10 are taken together with the carbon atom to which they are
attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-3 heteroatoms selected from O, N,
SO.sub.2, and S, said ring system optionally substituted with a
bond to the linking group or a bond to the chelator; and [0547]
R.sup.12 is independently C.sub.1-20 alkyl; [0548] R.sup.27 is
.dbd.O, C1-4 alkyl, or phenyl substituted with R.sup.28; [0549]
R.sup.28 is a phenoxy group substituted with 0-2 OCH.sub.3 groups;
[0550] R.sup.29 and R.sup.30 taken together with the nitrogen atom
through which they are attached form a C5-7 atom saturated ring
system substituted with R.sup.31; and [0551] R.sup.31 is a
benzyloxy group substituted with C1-4 alkyl. (9). A diagnostic
agent according to any one of embodiments 1-8 wherein the targeting
moiety is a matrix metalloproteinase inhibitor of the formulae (Ia)
or (Ib):
##STR00047##
[0551] wherein, [0552] R is OH; [0553] R.sup.1 is independently
selected at each occurrence from the group: H, OH, C.sub.1-3 alkyl,
C.sub.2-3 alkenyl, C.sub.2-3 alkynyl, and
heterocycle-S--CH.sub.2--; [0554] R.sup.2 is independently
C.sub.1-6 alkyl; [0555] X is C.dbd.O; [0556] R.sup.4 is
independently selected at each occurrence from the group: C.sub.1-6
alkyl, phenyl, and benzyl; [0557] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to the linking group or a
bond to the chelator; [0558] R.sup.6 is independently aryloxy
substituted with 0-3 R.sup.7; [0559] R.sup.7 is independently
halogen or methoxy; [0560] or alternatively, [0561] R.sup.1 and
R.sup.4 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally
substituted with a bond to the linking group or a bond to the
chelator; [0562] or alternatively, [0563] R.sup.1 and R.sup.2 may
be taken together to form a bridging group of the formula
--(CH.sub.2).sub.3--NH--, optionally substituted with a bond to the
linking group or a bond to the chelator; or [0564] R.sup.1 and
R.sup.2 taken together with the nitrogen and carbon atom through
which they are attached form a C.sub.5-7 atom saturated ring system
substituted with one or more substituents selected from the group
consisting of: a bond to Ln, a bond to Ch, and
--C(.dbd.O)--NR.sup.29R.sup.30; [0565] R.sup.8 is OH; [0566]
R.sup.9 and R.sup.9' are independently H, C.sub.1-6 alkyl
optionally substituted with a bond to the linking group or a bond
to the chelator, or are taken together with the carbon atom to
which R.sup.9 and R.sup.9' are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-1 heteroatoms selected from O, N, said ring system optionally
substituted with a bond to the linking group or a bond to the
chelator; [0567] R.sup.10 and R.sup.11 are independently H, or
C.sub.1-6 alkyl optionally substituted with a bond to the linking
group or a bond to the chelator, or are taken together with the
nitrogen atom to which they are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-1 heteroatoms selected from O, N, said ring system optionally
substituted with 0-3 R.sup.27, a bond to the linking group or a
bond to the chelator; [0568] or alternatively, [0569] R.sup.9 and
R.sup.10 are taken together with the carbon atom to which they are
attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-1 heteroatoms selected from O, N,
said ring system optionally substituted with a bond to the linking
group or a bond to the chelator; and [0570] R.sup.12 is
independently C.sub.1-6 alkyl; [0571] R.sup.27 is .dbd.O, C1-4
alkyl, or phenyl substituted with R.sup.28; [0572] R.sup.28 is a
phenoxy group substituted with 0-2 OCH.sub.3 groups; [0573]
R.sup.29 and R.sup.30 taken together with the nitrogen atom through
which they are attached form a C5-7 atom saturated ring system
substituted with R.sup.31; and [0574] R.sup.31 is a benzyloxy group
substituted with C1-4 alkyl. (10). A diagnostic agent according to
any one of embodiments 1-9 wherein the targeting moiety is a matrix
metalloproteinase inhibitor of the formulae (Ia) or (Ib): wherein:
[0575] R is --OH; [0576] R.sup.2 is C.sub.1-6 alkyl; [0577] X is
C.dbd.O; [0578] R.sup.3 is
[0578] ##STR00048## [0579] R.sup.1 and R.sup.4 are taken together
to form a bridging group of formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.2--; [0580] R.sup.5 is
NH(C1-6alkyl), substituted with a bond to the linking group or a
bond to the chelator. (11) A diagnostic agent according to any one
of embodiments 1-10, wherein:
R is --OH;
[0581] R.sup.9 is C.sub.1 alkyl substituted with a bond to Ln;
R.sup.10 and R.sup.11 taken together with the nitrogen atom to
which they are attached form a 5 atom saturated ring system, said
right system is substituted with 0-3 R.sup.27; R.sup.27 is .dbd.O,
C1-4 alkyl, or phenyl substituted with R.sup.28; and R.sup.28 is a
phenoxy group substituted with 0-2 OCH.sub.3 groups. (12) A
diagnostic agent according to any one of embodiments 1-11 wherein
the
R is --OH;
[0582] R.sup.1 and R.sup.2 taken together with the nitrogen and
carbon atom through which they are attached form a C.sub.5-7 atom
saturated ring system substituted with one or more substituents
selected from the group consisting of: a bond to Ln, a bond to Ch,
and --C(.dbd.O)--NR.sup.29R.sup.30; R.sup.29 and R.sup.30 taken
together with the nitrogen atom through which they are attached
form a C5-7 atom saturated ring system substituted with R.sup.31;
and R.sup.31 is a benzyloxy group substituted with C1-4 alkyl. (13)
A diagnostic agent according to any one of embodiments 1-12 wherein
the linking group is of the formula:
((W.sup.1).sub.h-(CR.sup.13R.sup.14).sub.g).sub.x--(Z).sub.k--((CR.sup.1-
3aR.sup.14a).sub.g'-(W.sup.2).sub.h').sub.x'; [0583] W.sup.1 and
W.sup.2 are independently selected at each occurrence from the
group: O, S, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2, SO.sub.2NH,
--(OCH.sub.2CH.sub.2).sub.76-84, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0584] aa is
independently at each occurrence an amino acid; [0585] Z is
selected from the group: aryl substituted with 0-3 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.16; [0586] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, PO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-3 R.sup.16, aryl substituted with 0-3 R.sup.16,
benzyl substituted with 0-3 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-3 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to the chelator; [0587] R.sup.16 is independently
selected at each occurrence from the group: a bond to the chelator,
COOR.sup.17, C(.dbd.O)NHR.sup.17, NHC(.dbd.O)R.sup.17, OH,
NHR.sup.17, SO.sub.3H, PO.sub.3H, --OPO.sub.3H.sub.2, --OSO.sub.3H,
aryl substituted with 0-3 R.sup.17, C.sub.1-5 alkyl substituted
with 0-1 R.sup.18, C.sub.1-5 alkoxy substituted with 0-1 R.sup.18,
and a 5-10 membered heterocyclic ring system containing 1-4
heteroatoms independently selected from N, S, and O and substituted
with 0-3 R.sup.17; [0588] R.sup.17 is independently selected at
each occurrence from the group: H, alkyl substituted with 0-1
R.sup.18, aryl substituted with 0-1 R.sup.18, a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-1 R.sup.18,
C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.18, polyalkylene
glycol substituted with 0-1 R.sup.18, carbohydrate substituted with
0-1 R.sup.18, cyclodextrin substituted with 0-1 R.sup.18, amino
acid substituted with 0-1 R.sup.18, polycarboxyalkyl substituted
with 0-1 R.sup.18, polyazaalkyl substituted with 0-1 R.sup.18,
peptide substituted with 0-1 R.sup.18, wherein the peptide is
comprised of 2-10 amino acids, 3,6-O-disulfo-B-D-galactopyranosyl,
bis(phosphonomethyl)glycine, and a bond to the chelator; [0589]
R.sup.18 is a bond to the chelator; [0590] k is selected from 0, 1,
and 2; [0591] h is selected from 0, 1, and 2; [0592] h' is selected
from 0, 1, and 2; [0593] g is selected from 0, 1, 2, 3, 4, 5, 6, 7,
8, 9, and 10; [0594] g' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8,
9, and 10; [0595] s is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,
and 10; [0596] s' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,
and 10; [0597] s'' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9,
and 10; [0598] t is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and
10; [0599] t' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and
10; [0600] x is selected from 0, 1, 2, 3, 4, and 5; and [0601] x'
is selected from 0, 1, 2, 3, 4, and 5. (14) A diagnostic agent
according to any one of embodiments 1-13 wherein [0602] W.sup.1 and
W.sup.2 are independently selected at each occurrence from the
group: O, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2,
--(CH.sub.2CH.sub.2O).sub.76-84--, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0603] aa is
independently at each occurrence an amino acid; [0604] Z is
selected from the group: aryl substituted with 0-1 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-1
R.sup.16; [0605] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-1 R.sup.16, aryl substituted with 0-1 R.sup.16,
benzyl substituted with 0-1 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-1 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to the chelator; [0606] k is 0 or 1; [0607] s is
selected from 0, 1, 2, 3, 4, and 5; [0608] s' is selected from 0,
1, 2, 3, 4, and 5; [0609] s'' is selected from 0, 1, 2, 3, 4, and
5; and [0610] t is selected from 0, 1, 2, 3, 4, and 5. (15) A
diagnostic agent according to embodiment 13 wherein wherein: [0611]
W.sup.1 is C(.dbd.O)NR.sup.15; [0612] h is 1; [0613] g is 3; [0614]
R.sup.13 and R.sup.14 are independently H; [0615] x is 1; [0616] k
is 0; [0617] g' is 0: [0618] h' is 1; [0619] W.sup.2 is NH; and
[0620] x' is 1. (16) A diagnostic agent according to embodiment 13
wherein [0621] x is 0; [0622] k is 1; [0623] Z is aryl substituted
with 0-3 R.sup.16; [0624] g' is 1; [0625] W.sup.2 is NH; [0626]
R.sup.13a and R.sup.14a are independently H; [0627] h' is 1; and
[0628] x' is 1. (17) A diagnostic agent according to embodiment 13
wherein [0629] W.sup.1 is C(.dbd.O)NR.sup.15; [0630] h is 1; [0631]
g is 2; [0632] R.sup.13 and R.sup.14 are independently H; [0633] x
is 1; [0634] k is 0; [0635] g' is 1; [0636] R.sup.13a and R.sup.14a
are independently H; or C1-5 alkyl substituted [0637] with 0-3
R.sup.16; [0638] R.sup.16 is SO.sub.3H; [0639] W.sup.2 is
NHC(.dbd.O) or NH; [0640] h' is 1; and [0641] x' is 2. (18). A
diagnostic agent according to embodiment 13 wherein [0642] W.sup.1
is C(O)NH; [0643] h is 1; [0644] g is 3; [0645] R.sup.13 and
R.sup.14 are independently H; [0646] k is 0; [0647] g' is 0; [0648]
x is 1; [0649] W.sup.2 is --NH(C.dbd.O)-- or
--(OCH.sub.2CH.sub.2).sub.76-84--; [0650] h' is 2; and [0651] x' is
1. (19) A diagnostic agent according to embodiment 13 wherein
[0652] x is 0; [0653] k is 0; [0654] g' is 3; [0655] h' is 1;
[0656] W.sup.2 is NH; and [0657] x' is 1. (20) A diagnostic agent
according to embodiment 13 wherein [0658] x is 0; [0659] Z is aryl
substituted with 0-3 R.sup.16, [0660] k is 1; [0661] g' is 1;
[0662] R.sup.13aR.sup.14a are independently H; [0663] W.sup.2 is
NHC(.dbd.O) or --(OCH2CH2).sub.76-84-; and [0664] x' is 1. (21) A
diagnostic agent according to embodiment 13 wherein [0665] W.sup.1
is C.dbd.O; [0666] g is 2; [0667] R.sup.13 and R.sup.14 are
independently H; [0668] k is 0; [0669] g' is 0; [0670] h' is 1;
[0671] W.sup.2 is NH; and [0672] x' is 1. (22) A compound according
to embodiment 1 wherein the linking group is absent. (23) A
diagnostic agent according to any one of embodiments 1-22 wherein
the chelator is a metal bonding unit having a formula selected from
the group:
[0672] ##STR00049## [0673] A.sup.1, A.sup.2, A.sup.3, A.sup.4,
A.sup.5, A.sup.6, A.sup.7, and A.sup.8 are independently selected
at each occurrence from the group: N, NR.sup.26, NR.sup.19,
NR.sup.19R.sup.20, S, SH, --S(Pg), O, OH, PR.sup.19,
PR.sup.19R.sup.20, --O--P(O)(R.sup.21)--O--, P(O)R.sup.21R.sup.22,
a bond to the targeting moiety and a bond to the linking group;
[0674] Pg is a thiol protecting group; [0675] E.sup.1, E.sup.2,
E.sup.3, E.sup.4, E.sup.5, E.sup.6, E.sup.7, and E.sup.8 are
independently a bond, CH, or a spacer group independently selected
at each occurrence from the group: C.sub.1-C.sub.16 alkyl
substituted with 0-3 R.sup.23, aryl substituted with 0-3 R.sup.23,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.23,
heterocyclo-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, wherein
the heterocyclo group is a 5-10 membered heterocyclic ring system
containing 1-4 heteroatoms independently selected from N, S, and O,
C.sub.6-10 aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-10 alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, and
a 5-10 membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23; [0676] R.sup.19 and R.sup.20 are each independently
selected from the group: a bond to the linking group, a bond to the
targeting moiety, hydrogen, C.sub.1-C.sub.10 alkyl substituted with
0-3 R.sup.23, aryl substituted with 0-3 R.sup.23, C.sub.1-10
cycloalkyl substituted with 0-3 R.sup.23, heterocyclo-C.sub.1-10
alkyl substituted with 0-3 R.sup.23, wherein the heterocyclo group
is a 5-10 membered heterocyclic ring system containing 1-4
heteroatoms independently selected from N, S, and O, C.sub.6-10
aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, C.sub.1-10
alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23, and an electron, provided that when one of R.sup.19 or
R.sup.20 is an electron, then the other is also an electron; [0677]
R.sup.21 and R.sup.22 are each independently selected from the
group: a bond to the linking group, a bond to the targeting moiety,
--OH, C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23, aryl
substituted with 0-3 R.sup.23, C.sub.3-10 cycloalkyl substituted
with 0-3 R.sup.23, heterocyclo-C.sub.1-10 alkyl substituted with
0-3 R.sup.23, wherein the heterocyclo group is a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O, C.sub.6-10 aryl-C.sub.1-10 alkyl
substituted with 0-3 R.sup.23, C.sub.1-10 alkyl-C.sub.6-10
aryl-substituted with 0-3 R.sup.23, and a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-3 R.sup.23 [0678]
R.sup.23 is independently selected at each occurrence from the
group: a bond to the linking group, a bond to the targeting moiety,
.dbd.O, F, Cl, Br, I, --CF.sub.3, --CN, --CO.sub.2R.sup.24,
--C(.dbd.O)R.sup.24, --C(.dbd.O)N(R.sup.24).sub.2, --CHO,
--CH.sub.2OR.sup.24, --OC(.dbd.O)R.sup.24, --OC(.dbd.O)OR.sup.24a,
--OR.sup.24, --OC(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25C(.dbd.O)R.sup.24, --NR.sup.25C(.dbd.O)OR.sup.24a,
--NR.sup.25C(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25SO.sub.2N(R.sup.24).sub.2, --NR.sup.25SO.sub.2R.sup.24a,
--SO.sub.3H, --SO.sub.2R.sup.24a, --SR.sup.24,
--S(.dbd.O)R.sup.24a, --SO.sub.2N(R.sup.24).sub.2,
--N(R.sup.24).sub.2, --NHC(.dbd.S)NHR.sup.24, .dbd.NOR.sup.24,
NO.sub.2, --C(.dbd.O)NHOR.sup.24, --C(.dbd.O)NHNR.sup.24R.sup.24a,
--OCH.sub.2CO.sub.2H, 2-(1-morpholino)ethoxy, C.sub.1-C.sub.5
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 cycloalkylmethyl, C.sub.2-C.sub.6 alkoxyalkyl, aryl
substituted with 0-2 R.sup.24, and a 5-10 membered heterocyclic
ring system containing 1-4 heteroatoms independently selected from
N, S, and O; and [0679] wherein at least one of A.sup.1, A.sup.2,
A.sup.3, A.sup.4, A.sup.5, A.sup.6, A.sup.7, A.sup.8 or R.sup.23 is
a bond to the linking group or targeting moiety; [0680] R.sup.24,
R.sup.24a, and R.sup.25 are independently selected at each
occurrence from the group: a bond to the linking group, a bond to
the targeting moiety, H, C.sub.1-C.sub.6 alkyl, phenyl, benzyl,
C.sub.1-C.sub.6 alkoxy, halide, nitro, cyano, and trifluoromethyl;
and [0681] R.sup.26 is a co-ordinate bond to a metal or a hydrazine
protecting group; or a pharmaceutically acceptable salt thereof.
(24) A diagnostic agent according to any one of embodiments 1-23
wherein: [0682] A.sup.1, A.sup.2, A.sup.3, A.sup.4, A.sup.5,
A.sup.6, A.sup.7, and A.sup.8 are independently selected at each
occurrence from the group: NR.sup.19, NR.sup.19R.sup.20, S, SH, OH,
a bond to the targeting moiety and a bond to the linking group;
[0683] E.sup.1, E.sup.2, E.sup.3, E.sup.4, E.sup.5, E.sup.6,
E.sup.7, and E.sup.8 are independently a bond, CH, or a spacer
group independently selected at each occurrence from the group:
C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23, aryl
substituted with 0-3 R.sup.23, C.sub.3-10 cycloalkyl substituted
with 0-3 R.sup.23, and a 5-10 membered heterocyclic ring system
containing 1-4 heteroatoms independently selected from N, S, and O
and substituted with 0-3 R.sup.23; [0684] wherein at least one of
A.sup.1, A.sup.2, A.sup.3, A.sup.4, A.sup.5, A.sup.6, A.sup.7,
A.sup.8 and R.sup.23 is a bond to the linking group or the
targeting moiety; [0685] R.sup.19, and R.sup.20 are each
independently selected from the group: a bond to the targeting
moiety, a bond to the linking group, hydrogen, C.sub.1-C.sub.10
alkyl substituted with 0-3 R.sup.23, aryl substituted with 0-3
R.sup.23, a 5-10 membered heterocyclic ring system containing 1-4
heteroatoms independently selected from N, S, and O and substituted
with 0-3 R.sup.23, and an electron, provided that when one of
R.sup.19 or R.sup.20 is an electron, then the other is also an
electron; [0686] R.sup.23 is independently selected at each
occurrence from the group: a bond to the targeting moiety, a bond
to the linking group, .dbd.O, F, Cl, Br, I, --CF.sub.3, --CN,
--CO.sub.2R.sup.24, --C(.dbd.O)R.sup.24,
--C(.dbd.O)N(R.sup.24).sub.2, --CH.sub.2OR.sup.24,
--OC(.dbd.O)R.sup.24, --OC(.dbd.O)OR.sup.24a, --OR.sup.24,
--OC(.dbd.O)N(R.sup.24).sub.2, --NR.sup.25C(.dbd.O)R.sup.24,
--NR.sup.25C(.dbd.O)OR.sup.24a,
--NR.sup.25C(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25SO.sub.2N(R.sup.24).sub.2, --NR.sup.25SO.sub.2R.sup.24a,
--SO.sub.3H, --SO.sub.2R.sup.24a, --S(.dbd.O)R.sup.24a,
--SO.sub.2N(R.sup.24).sub.2, --N(R.sup.24).sub.2,
--NHC(.dbd.S)NHR.sup.24, .dbd.NOR.sup.18,
--C(.dbd.O)NHNR.sup.18R.sup.18a, --OCH.sub.2CO.sub.2H, and
2-(1-morpholino)ethoxy; and [0687] R.sup.24, R.sup.24a, and
R.sup.25 are independently selected at each occurrence from the
group: a bond to the linking group, H, and C.sub.1-C.sub.6 alkyl.
(25) A diagnostic agent according to any one of embodiments 1-24
wherein the chelator is of the formula:
[0687] ##STR00050## [0688] A.sup.1 is a bond to the linking group;
[0689] A.sup.2, A.sup.4, and A.sup.6 are each N; [0690] A.sup.3,
A.sup.5, A.sup.7 and A.sup.8 are each OH; [0691] E.sup.1, E.sup.2,
and E.sup.4 are C2 alkyl; [0692] E.sup.3, E.sup.5, E.sup.7, and
E.sup.8 are C.sub.2 alkyl substituted with 0-1 R.sup.23; [0693]
R.sup.23 is .dbd.O. (26) A diagnostic agent according to any one of
embodiments 1-25 wherein the chelator is of the formula: [0694]
C.sub.h is
##STR00051##
[0694] wherein: [0695] A5 is a bond to Ln; [0696] A.sup.1, A.sup.3,
A.sup.7 and A.sup.8 are each OH; [0697] A.sup.2, A.sup.4 and
A.sup.6 are each NH; [0698] E.sup.1, E.sup.3, E.sup.5, E.sup.7, and
E.sup.8 are C.sub.2 alkyl substituted with 0-1 R.sup.23; [0699]
E.sup.2, and E.sup.4, are C.sub.2 alkyl; [0700] R.sup.23 is .dbd.O.
(27) A diagnostic agent according to any one of embodiments 1-26
wherein the chelator is of the formula:
[0700] ##STR00052## [0701] A.sup.1, A.sup.2, A.sup.3 and A.sup.4
are each N; [0702] A.sup.5, A.sup.6 and A.sup.8 are each OH; [0703]
A.sup.7 is a bond to L.sub.n; [0704] E.sup.1, E.sup.2, E.sup.3,
E.sup.4 are each independently C.sub.2 alkyl; and [0705] E.sup.5,
E.sup.6, E.sup.7, E.sup.8 are each independently C.sub.2 alkyl
substituted with 0-1 R.sup.23; [0706] R.sup.23 is .dbd.O. (28) A
diagnostic agent according to any one of embodiments 1-27 wherein
the chelator is of the formula:
[0706] ##STR00053## [0707] A.sup.1 is NR.sup.26; [0708] R.sup.26 is
a co-ordinate bond to a metal or a hydrazine protecting group;
[0709] E.sup.1 is a bond; [0710] A.sup.2 is NHR.sup.19; [0711]
R.sup.19 is a heterocycle substituted with R.sup.23, the
heterocycle being selected from pyridine and pyrimidine; [0712]
R.sup.23 is selected from a bond to the linking group,
C(.dbd.O)NHR.sup.24 and C(.dbd.O)R.sup.24; and [0713] R.sup.24 is a
bond to the linking group. (29) A diagnostic agent according to any
one of embodiments 1-28 wherein the chelator is of the formula:
##STR00054##
[0713] wherein: [0714] A.sup.1 and A.sup.5 are each --S(Pg); [0715]
Pg is a thiol protecting group; [0716] E.sup.1 and E.sup.4 are
C.sub.2 alkyl substituted with 0-1 R.sup.23; [0717] R.sup.23 is
.dbd.O; [0718] A.sup.2 and A.sup.4 are each --NH; [0719] E.sup.2 is
CH.sub.2; [0720] E.sup.3 is C.sub.1-3 alkyl substituted with 0-1
R.sup.23; [0721] A.sup.3 is a bond to Ln. (30) A diagnostic agent
according to any one of embodiments 1-29 wherein the chelator is of
the formula:
##STR00055##
[0721] wherein: [0722] A.sup.1 is a bond to Ln; [0723] E.sup.1 is
C.sub.1 alkyl substituted by R.sup.23; [0724] A.sup.2 is NH; [0725]
E.sup.2 is C.sub.2 alkyl substituted with 0-1R.sup.23; [0726]
A.sup.3 is --O--P(O)(R.sup.21)--O; [0727] E.sup.3 is C.sub.1 alkyl;
[0728] A.sup.4 and A.sup.5 are each --O--; [0729] E.sup.4 and
E.sup.6 are each independently C.sub.1-16 alkyl substituted with
0-1R.sup.23; [0730] E.sup.5 is C.sub.1 alkyl; [0731] R.sup.21 is
--OH; and [0732] R.sup.23 is .dbd.O. (31) A diagnostic agent
according to embodiment 1 having the formula:
[0732] (Q).sub.d-L.sub.n-C.sub.h
wherein, Q is a compound of Formulae (Ia) or (Ib):
##STR00056##
wherein, [0733] R is independently OH or --CH.sub.2SH; [0734]
R.sup.1 is independently selected at each occurrence from the
group: H, OH, C.sub.1-3 alkyl, C.sub.2-3 alkenyl, C.sub.2-3
alkynyl, and heterocycle-S--CH.sub.2--; [0735] R.sup.2 is
independently C.sub.1-20 alkyl; [0736] X is independently C.dbd.O
or SO.sub.2, provided when X is C.dbd.O, R.sup.3 is
##STR00057##
[0736] and when X is SO.sub.2, R.sup.3 is independently selected
from the group: aryl substituted with 0-2 R.sup.6, and heterocycle
substituted with 0-2 R.sup.6; [0737] R.sup.4 is independently
selected at each occurrence from the group: C.sub.1-6 alkyl,
phenyl, and benzyl; [0738] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to L.sub.n; [0739] R.sup.6
is independently aryloxy substituted with 0-3 R.sup.7; [0740]
R.sup.7 is independently halogen or methoxy; [0741] or
alternatively, [0742] R.sup.1 and R.sup.4 may be taken together to
form a bridging group of the formula
--(CH).sub.3--O-phenyl-CH.sub.2--, optionally substituted with a
bond to L.sub.n; [0743] or alternatively, [0744] R.sup.1 and
R.sup.2 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--NH--, optionally substituted with a
bond to L.sub.n; or [0745] R.sup.1 and R.sup.2 taken together with
the nitrogen and carbon atom through which they are attached form a
C.sub.5-7 atom saturated ring system substituted with one or more
substituents selected from the group consisting of: a bond to Ln, a
bond to Ch, and --C(.dbd.O)--NR.sup.29R.sup.30; [0746] R.sup.8 is
independently at each occurrence OH or phenyl, optionally
substituted with a bond to L.sub.n, provided that when R.sup.8 is
phenyl, R.sup.10 is --C(.dbd.O)--CR.sup.12--NH--CH(CH.sub.3)--COOH;
[0747] R.sup.9 and R.sup.9' are independently H, C.sub.1-6 alkyl
optionally substituted with a bond to L.sub.n, or are taken
together with the carbon atom to which they are attached to form a
5-7 atom saturated, partially unsaturated or aromatic ring system
containing 0-3 heteroatoms selected from O, N, SO.sub.2, and S,
said ring system substituted with R.sup.6 and optionally
substituted with a bond to L.sub.n; [0748] R.sup.10 and R.sup.11
are independently H, or C.sub.1-6 alkyl optionally substituted with
a bond to L.sub.n, or are taken together with the nitrogen atom to
which they are attached to form a 5-7 atom saturated, partially
unsaturated or aromatic ring system containing 0-3 heteroatoms
selected from O, N, SO.sub.2 and S, said ring system optionally
substituted with 0-3 R.sup.27 or a bond to L.sub.n; [0749] or
alternatively, [0750] R.sup.9 and R.sup.10 are taken together with
the carbon atom to which they are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-3 heteroatoms selected from O, N, SO.sub.2 and S, said ring
system optionally substituted with a bond to L.sub.n; [0751]
R.sup.12 is independently C.sub.1-20 alkyl; [0752] d is selected
from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; [0753] L.sub.n is a linking
group having the formula:
[0753]
((W.sup.1).sub.h-(CR.sup.13R.sup.14).sub.g).sub.x--(Z).sub.k--((C-
R.sup.13aR.sup.14a).sub.g'-(W.sup.2).sub.h').sub.x'; [0754] W.sup.1
and W.sup.2 are independently selected at each occurrence from the
group: O, S, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2, SO.sub.2NH,
--(OCH.sub.2CH.sub.2).sub.76-84, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t', [0755] aa is
independently at each occurrence an amino acid; [0756] Z is
selected from the group: aryl substituted with 0-3 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.16; [0757] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, PO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-3 R.sup.16, aryl substituted with 0-3 R.sup.16,
benzyl substituted with 0-3 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-3 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to C.sub.h; [0758] R.sup.16 is independently selected at
each occurrence from the group: a bond to C.sub.h, COOR.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)R.sup.17, OH, NHR.sup.17,
SO.sub.3H, PO.sub.3H, --OPO.sub.3H.sub.2, --OSO.sub.3H, aryl
substituted with 0-3 R.sup.17, C.sub.1-5 alkyl substituted with 0-1
R.sup.18, C.sub.1-5 alkoxy substituted with 0-1 R.sup.18, and a
5-10 membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.17; [0759] R.sup.17 is independently selected at each
occurrence from the group: H, alkyl substituted with 0-1 R.sup.18,
aryl substituted with 0-1 R.sup.18, a 5-10 membered heterocyclic
ring system containing 1-4 heteroatoms independently selected from
N, S, and O and substituted with 0-1 R.sup.18, C.sub.3-10
cycloalkyl substituted with 0-1 R.sup.18, polyalkylene glycol
substituted with 0-1 R.sup.18, carbohydrate substituted with 0-1
R.sup.18, cyclodextrin substituted with 0-1 R.sup.18, amino acid
substituted with 0-1 R.sup.18, polycarboxyalkyl substituted with
0-1 R.sup.18, polyazaalkyl substituted with 0-1 R.sup.18, peptide
substituted with 0-1 R.sup.18, wherein the peptide is comprised of
2-10 amino acids, 3,6-O-disulfo-B-D-galactopyranosyl,
bis(phosphonomethyl)glycine, and a bond to C.sub.h; [0760] R.sup.18
is a bond to C.sub.h; [0761] k is selected from 0, 1, and 2; [0762]
h is selected from 0, 1, and 2; [0763] h' is selected from 0, 1,
and 2; [0764] g is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and
10; [0765] g' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and
10; [0766] s is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0767] s' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0768] s'' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0769] t is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0770] t' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0771] x is selected from 0, 1, 2, 3, 4, and 5; [0772] x' is
selected from 0, 1, 2, 3, 4, and 5; [0773] C.sub.h is a metal
bonding unit having a formula selected from the group:
[0773] ##STR00058## [0774] A.sup.1, A.sup.2, A.sup.3, A.sup.4,
A.sup.5, A.sup.6, A.sup.7, and A.sup.8 are independently selected
at each occurrence from the group: N, NR.sup.26, NR.sup.19,
NR.sup.19R.sup.20, S, SH, --S(Pg), O, OH, PR.sup.19,
PR.sup.19R.sup.20, --O--P(O)(R.sup.21)--O--, P(O)R.sup.21R.sup.22,
a bond to the targeting moiety and a bond to the linking group;
[0775] Pg is a thiol protecting group; [0776] E.sup.1, E.sup.2,
E.sup.3, E.sup.4, E.sup.5, E.sup.6, E.sup.7, and E.sup.8 are
independently a bond, CH, or a spacer group independently selected
at each occurrence from the group: C.sub.1-C.sub.16 alkyl
substituted with 0-3 R.sup.23, aryl substituted with 0-3 R.sup.23,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.23,
heterocyclo-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, wherein
the heterocyclo group is a 5-10 membered heterocyclic ring system
containing 1-4 heteroatoms independently selected from N, S, and O,
C.sub.6-10 aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-10 alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, and
a 5-10 membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23; [0777] R.sup.19 and R.sup.20 are each independently
selected from the group: a bond to the linking group, a bond to the
targeting moiety, hydrogen, C.sub.1-C.sub.10 alkyl substituted with
0-3 R.sup.23, aryl substituted with 0-3 R.sup.23, C.sub.1-10
cycloalkyl substituted with 0-3 R.sup.23, heterocyclo-C.sub.1-10
alkyl substituted with 0-3 R.sup.23, wherein the heterocyclo group
is a 5-10 membered heterocyclic ring system containing 1-4
heteroatoms independently selected from N, S, and O, C.sub.6-10
aryl-C.sub.1-10 alkyl substituted with 0-3 R.sup.23, C.sub.1-10
alkyl-C.sub.6-10 aryl-substituted with 0-3 R.sup.23, a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.23, and an electron, provided that when one of R.sup.19 or
R.sup.20 is an electron, then the other is also an electron; [0778]
R.sup.21 and R.sup.22 are each independently selected from the
group: a bond to the linking group, a bond to the targeting moiety,
--OH, C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23,
C.sub.1-C.sub.10 alkyl substituted with 0-3 R.sup.23, aryl
substituted with 0-3 R.sup.23, C.sub.3-10 cycloalkyl substituted
with 0-3 R.sup.23, heterocyclo-C.sub.1-10 alkyl substituted with
0-3 R.sup.23, wherein the heterocyclo group is a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O, C.sub.6-10 aryl-C.sub.1-10 alkyl
substituted with 0-3 R.sup.23, C.sub.1-10 alkyl-C.sub.6-10
aryl-substituted with 0-3 R.sup.23, and a 5-10 membered
heterocyclic ring system containing 1-4 heteroatoms independently
selected from N, S, and O and substituted with 0-3 R.sup.23; [0779]
R.sup.23 is independently selected at each occurrence from the
group: a bond to the linking group, a bond to the targeting moiety,
.dbd.O, F, Cl, Br, I, --CF.sub.3, --CN, --CO.sub.2R.sup.24,
--C(.dbd.O)R.sup.24, --C(.dbd.O)N(R.sup.24).sub.2, --CHO,
--CH.sub.2OR.sup.24, --OC(.dbd.O)R.sup.24, --OC(.dbd.O)OR.sup.24a,
--OR.sup.24, --OC(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25C(.dbd.O)R.sup.24, --NR.sup.25C(.dbd.O)OR.sup.24a,
--NR.sup.25C(.dbd.O)N(R.sup.24).sub.2,
--NR.sup.25SO.sub.2N(R.sup.24).sub.2, --NR.sup.25SO.sub.2R.sup.24a,
--SO.sub.3H, --SO.sub.2R.sup.24a, --SR.sup.24,
--S(.dbd.O)R.sup.24a, --SO.sub.2N(R.sup.24).sub.2,
--N(R.sup.24).sub.2, --NHC(.dbd.S)NHR.sup.24, .dbd.NOR.sup.24,
NO.sub.2, --C(.dbd.O)NHOR.sup.24, --C(.dbd.O)NHNR.sup.24R.sup.24a,
--OCH.sub.2CO.sub.2H, 2-(1-morpholino) ethoxy, C.sub.1-C.sub.5
alkyl, C.sub.2-C.sub.4 alkenyl, C.sub.3-C.sub.6 cycloalkyl,
C.sub.3-C.sub.6 cycloalkylmethyl, C.sub.2-C.sub.6 alkoxyalkyl, aryl
substituted with 0-2 R.sup.24, and a 5-10 membered heterocyclic
ring system containing 1-4 heteroatoms independently selected from
N, S, and O; and [0780] wherein at least one of A.sup.1, A.sup.2,
A.sup.3, A.sup.4, A.sup.5, A.sup.6, A.sup.7, A.sup.8 or R.sup.23 is
a bond to the linking group or targeting moiety; [0781] R.sup.24,
R.sup.24a, and R.sup.25 are independently selected at each
occurrence from the group: a bond to the linking group, a bond to
the targeting moiety, H, C.sub.1-C.sub.6 alkyl, phenyl, benzyl,
C.sub.1-C.sub.6 alkoxy, halide, nitro, cyano, and trifluoromethyl;
and [0782] R.sup.26 is a co-ordinate bond to a metal or a hydrazine
protecting group; or [0783] a pharmaceutically acceptable salt
thereof. (32) A diagnostic agent according to Embodiment 31,
wherein: [0784] h' is 1; [0785] W.sup.2 is NH; and [0786] x' is 1.
(33) A diagnostic agent according to any one of embodiments 1,
wherein: [0787] x is 0; [0788] Z is aryl substituted with 0-3
R.sup.16; [0789] k is 1; [0790] g' is 1; [0791] R.sup.13aR.sup.14a
are independently H; [0792] W.sup.2 is NHC(.dbd.O) or
--(OCH2CH2).sub.76-84-; and [0793] x' is 1. (34) A diagnostic agent
according to any one of embodiments 31-33, wherein: [0794] W.sup.1
is C.dbd.O; [0795] g is 2; [0796] R.sup.13 and R.sup.14 are
independently H; [0797] k is 0; [0798] g' is 0; [0799] h' is 1;
[0800] W.sup.2 is NH; and [0801] x' is 1. (35) A diagnostic agent
according to any one of embodiment 31-34, wherein: [0802]
2-{[5-(3-{2-[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicyclo[10.-
2.2]hexadeca-1(15),12(16),13-triene-10-carbonyl)-amino]-acetylamino}-propy-
lcarbamoyl)-pyridin-2-yl]-hydrazonomethyl}-benzenesulfonic acid;
[0803]
2-{[5-(4-{[(6-Hydroxycarbamoyl-7-isobutyl-8-oxo-2-oxa-9-aza-bicyclo[10.2.-
2]hexadeca-1(15),12(16),13-triene-10-carbonyl)-amino]-methyl}-benzylcarbam-
oyl)-pyridin-2-yl]-hydrazonomethyl}-benzenesulfonic acid; [0804]
2-[7-({N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl-
)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonyla-
mino}acetylamino)propyl]carbamoyl}methyl)-1,4,7,10-tetraaza-4,10-bis(carbo-
xymethyl)cyclododecyl]acetic acid; [0805]
2-{7-[(N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-carbonyla-
mino}methyl)phenyl]methyl}carbamoyl)methyl]-1,4,7,10-tetraaza-4,10-bis(car-
boxymethyl)cyclododecyl}acetic acid; [0806]
2-(7-{[N-(1-(N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methyl-
propyl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]car-
bonylamino}acetylamino)propyl]carbamoyl}-2-sulfoethyl)carbamoyl]methyl}-1,-
4,7,10-tetraaza-4,10-bis(carboxymethyl)cyclododecyl)acetic acid;
[0807]
2-(7({N-[1-(N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpr-
opyl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-carb-
onylamino}methyl)phenyl]methyl}carbamoyl)-2-sulfoethyl]carbamoyl}methyl)-1-
,4,7,10-tetraaza-4,10-bis(carboxymethyl)cyclododecyl)acetic acid;
[0808]
2-({2-[({N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylprop-
yl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbony-
lamino}acetylamino)propyl]carbamoyl}methyl)(carboxymethyl)amino}ethyl){2-[-
bis(carboxymethyl)amino]ethyl}amino]acetic acid; [0809]
2-[(2-{[N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl-
)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-carbonyl-
amino}methyl)phenyl]methyl}carbamoyl)methyl](carboxymethyl)amino}ethyl){2--
[bis(carboxymethyl)amino]ethyl}amino]acetic acid; [0810]
N-[3-(2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-11-ox-
a-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylamino}ac-
etylamino)propyl]-4,5-bis[2-(ethoxyethylthio)acetylamino]pentanamide;
[0811]
N-{[4-({[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylpropyl)-
-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carbonylam-
ino}methyl)-phenyl]methyl}-4,5-bis[2-(ethoxyethylthio)acetylamino]-pentana-
mide; [0812]
1-(1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamino)-.alpha.,.omega.-dica-
rbonylPEG.sub.3400-2-{[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylp-
ropyl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]carb-
onylamino}-N-(3-aminopropyl)acetamide; [0813]
1-(1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamino)-.alpha.,.omega.-dica-
rbonylPEG.sub.3400-[7-(N-hydroxycarbamoyl)(3S,6R,7S)-4-aza-6-(2-methylprop-
yl)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-trien-3-yl]-N-{[4--
(aminomethyl)phenyl]methyl}carboxamide conjugate; [0814]
2-[2-({5-[N-(5-(N-hydroxycarbamoyl)(5R)-5-{3-[4-(3,4-dimethoxyphenoxy)phe-
nyl]-3-methyl-2-oxopyrrolidinyl}pentyl)carbamoyl](2-pyridyl)}amino)(1Z)-2--
azavinyl]benzenesulfonic acid; [0815]
2-(2-{[5-(N-{3-[3-(N-hydroxycarbamoyl)(4S)-4-({4-[(4-methylphenyl)methoxy-
]piperidyl}carbonyl)piperidyl]-3-oxopropyl}carbamoyl)(2-pyridyl)]amino}(1Z-
)-2-azavinyl)benzenesulfonic acid; and
##STR00059##
[0815] (36) A diagnostic agent according to any one of embodiments
31-35 wherein the diagnostic metal is selected from the group
consisting of: a paramagnetic metal, a ferromagnetic metal, a
gamma-emitting radioisotope, or an x-ray absorber. (37) A
diagnostic agent according to any one of embodiments 31-36 wherein
the diagnostic metal is radioisotope selected from the group
consisting of .sup.99mTc, .sup.95Tc, .sup.111In, .sup.62Cu,
.sup.64Cu, .sup.67Ga, and .sup.68Ga. (38). A diagnostic agent
according to any one of embodiments 31-37 further comprising a
first ancillary ligand and a second ancillary ligand capable of
stabilizing the radioisotope. (39) A diagnostic agent according to
Embodiment 37, wherein the radioisotope is .sup.99mTc. (40) A
diagnostic agent according to Embodiment 37, wherein the
radioisotope is .sup.111In. (41) A diagnostic agent according to
embodiment 36 wherein the paramagnetic metal ion is selected from
the group consisting of Gd(III), Dy(III), Fe(III), and Mn(II).
(42). A diagnostic agent according to embodiment 36 wherein the
x-ray absorber is a metal is selected from the group consisting of:
Re, Sm, Ho, Lu, Pm, Y, Bi, Pd, Gd, La, Au, Au, Yb, Dy, Cu, Rh, Ag,
and Ir. (43) A diagnostic composition comprising a compound
according to any one of embodiments 1-42 or a pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable carrier.
(44) A kit comprising a compound of to any one of embodiments 1-42,
or a pharmaceutically acceptable salt form thereof and a
pharmaceutically acceptable carrier. (45) A kit according to
Embodiment 44, wherein the kit further comprises one or more
ancillary ligands and a reducing agent. (46) A kit according to
Embodiment 45, wherein the ancillary ligands are tricine and TPPTS.
(47) A kit according to Embodiment 45, wherein the reducing agent
is tin(II). (48) A diagnostic agent comprising an echogenic gas and
a compound, wherein the compound comprises: [0816] i) 1-10
targeting moieties; [0817] ii) a surfactant (Sf); and [0818] iii)
0-1 linking groups between the targeting moiety and surfactant;
wherein the targeting moiety is a matrix metalloproteinase
inhibitor; and [0819] wherein the surfactant is capable of forming
an echogenic gas filled lipid sphere or microbubble. (49) A
diagnostic agent according to embodiment 48, wherein the targeting
moiety is a matrix metalloproteinase inhibitor having an inhibitory
constant K.sub.i of <1000 nM. (50) A diagnostic agent according
to any one of embodiments 48-49, wherein the targeting moiety is a
matrix metalloproteinase inhibitor having an inhibitory constant
K.sub.i of <100 nM. (51) A diagnostic agent according to
embodiment 48, comprising 1-5 targeting moieties. (52). A
diagnostic agent according to embodiment 48, comprising one
targeting moiety. (53) A diagnostic agent according to any one of
embodiments 48-52, wherein the targeting moiety is an inhibitor of
one or more matrix metalloproteinases selected from the group
consisting of MMP-1, MMP-2, MMP-3, MMP-9, and MMP-14. (54) A
diagnostic agent according to any one of embodiments 48-53, wherein
the targeting moiety is an inhibitor of one or more matrix
metalloproteinases selected from the group consisting of MMP-2,
MMP-9, and MMP-14. (55) A diagnostic agent according to embodiment
48, wherein the targeting moiety is of the formulae (Ia) or
(Ib):
##STR00060##
[0819] wherein, [0820] R is independently OH or --CH.sub.2SH;
[0821] R.sup.1 is independently selected at each occurrence from
the group: H, OH, C.sub.1-3 alkyl, C.sub.2-3 alkenyl, C.sub.2-3
alkynyl, and heterocycle-S--CH.sub.2--; [0822] R.sup.2 is
independently C.sub.1-20 alkyl; [0823] X is independently C.dbd.O
or SO.sub.2, provided when X is C.dbd.O, R.sup.3 is
##STR00061##
[0823] and when X is SO.sub.2, R.sup.3 is independently selected
from the group: aryl substituted with 0-2 R.sup.6, and heterocycle
substituted with 0-2 R.sup.6; [0824] R.sup.4 is independently
selected at each occurrence from the group: C.sub.1-6 alkyl,
phenyl, and benzyl; [0825] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to the linking group or a
bond to the surfactant; [0826] R.sup.6 is independently aryloxy
substituted with 0-3 R.sup.7; [0827] R.sup.7 is independently
halogen or methoxy; [0828] or alternatively, [0829] R.sup.1 and
R.sup.4 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally
substituted with a bond to the linking group or a bond to the
surfactant; [0830] or alternatively, [0831] R.sup.1 and R.sup.2 may
be taken together to form a bridging group of the formula
--(CH.sub.2).sub.3--NH--, optionally substituted with a bond to the
linking group or a bond to the surfactant; or [0832] R.sup.1 and
R.sup.2 taken together with the nitrogen and carbon atom through
which they are attached form a C.sub.5-7 atom saturated ring system
substituted with one or more substituents selected from the group
consisting of: a bond to Ln, a bond to Sf, and
--C(.dbd.O)--NR.sup.29R.sup.30; [0833] R.sup.8 is independently at
each occurrence OH or phenyl, optionally substituted with a bond to
the linking group or a bond to the surfactant, provided that when
R.sup.8 is phenyl, R.sup.10 is
--C(.dbd.O)--CR.sup.12--NH--CH(CH.sub.3)--COOH; [0834] R.sup.9 and
R.sup.9' are independently H, C.sub.1-6 alkyl optionally
substituted with a bond to the linking group or a bond to the
surfactant, or are taken together with the carbon atom to which
R.sup.9 and R.sup.9' are attached to form a 5-7 atom saturated,
partially unsaturated or aromatic ring system containing 0-3
heteroatoms selected from O, N, SO.sub.2 and S, said ring system
substituted with R.sup.6 and optionally substituted with a bond to
the linking group or a bond to the surfactant; [0835] R.sup.10 and
R.sup.11 are independently N, or C.sub.1-6 alkyl optionally
substituted with a bond to the linking group or a bond to the
surfactant, or are taken together with the nitrogen atom to which
they are attached to form a 5-7 atom saturated, partially
unsaturated or aromatic ring system containing 0-3 heteroatoms
selected from O, N, SO.sub.2 and S, said ring system optionally
substituted with 0-3 R.sup.27, a bond to the linking group or a
bond to the surfactant; [0836] or alternatively, [0837] R.sup.9 and
R.sup.10 are taken together with the carbon atom to which they are
attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-3 heteroatoms selected from O, N,
SO.sub.2 and S, said ring system optionally substituted with a bond
to the linking group or a bond to the surfactant; and [0838]
R.sup.12 is independently C.sub.1-20 alkyl; [0839] R.sup.27 is
.dbd.O, C.sub.1-4 alkyl, or phenyl substituted with R.sup.28;
[0840] R.sup.28 is a phenoxy group substituted with 0-2 OCH.sub.3
groups; [0841] R.sup.29 and R.sup.30 taken together with the
nitrogen atom through which they are attached form a C5-7 atom
saturated ring system substituted with R.sup.31; and [0842]
R.sup.31 is a benzyloxy group substituted with C.sub.1-4 alkyl.
(56) A diagnostic agent according to embodiment 55 wherein wherein
the targeting moiety is a matrix metalloproteinase inhibitor of the
formulae (Ia) or (Ib):
##STR00062##
[0842] wherein, [0843] R is OH; [0844] R.sup.1 is independently
selected at each occurrence from the group: H, OH, C.sub.1-3 alkyl,
C.sub.2-3 alkenyl, C.sub.2-3 alkynyl, and
heterocycle-S--CH.sub.2--; [0845] R.sup.2 is independently
C.sub.1-6 alkyl; [0846] X is C.dbd.O; [0847] R.sup.4 is
independently selected at each occurrence from the group: C.sub.1-6
alkyl, phenyl, and benzyl; [0848] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to the linking group or a
bond to the surfactant; [0849] R.sup.6 is independently aryloxy
substituted with 0-3 R.sup.7; [0850] R.sup.7 is independently
halogen or methoxy; [0851] or alternatively, [0852] R.sup.1 and
R.sup.4 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally
substituted with a bond to the linking group or a bond to the
surfactant; [0853] or alternatively, [0854] R.sup.1 and R.sup.2 may
be taken together to form a bridging group of the formula
--(CH.sub.2).sub.3--NH--, optionally substituted with a bond to the
linking group or a bond to the surfactant; or [0855] R.sup.1 and
R.sup.2 taken together with the nitrogen and carbon atom through
which they are attached form a C.sub.5-7 atom saturated ring system
substituted with one or more substituents selected from the group
consisting of: a bond to Ln, a bond to Sf, and
--C(.dbd.O)--NR.sup.29R.sup.30; [0856] R.sup.8 is OH; [0857]
R.sup.9 and R.sup.9' are independently H, C.sub.1-6 alkyl
optionally substituted with a bond to the linking group or a bond
to the surfactant, or are taken together with the carbon atom to
which R.sup.9 and R.sup.9' are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-1 heteroatoms selected from O, N, said ring system optionally
substituted with a bond to the linking group or a bond to the
surfactant; [0858] R.sup.10 and R.sup.11 are independently H, or
C.sub.1-6 alkyl optionally substituted with a bond to the linking
group or a bond to the surfactant, or are taken together with the
nitrogen atom to which they are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-1 heteroatoms selected from O, N, said ring system optionally
substituted with 0-3 R.sup.27, a bond to the linking group or a
bond to the surfactant; [0859] or alternatively, [0860] R.sup.9 and
R.sup.10 are taken together with the carbon atom to which they are
attached to form a 5-7 atom saturated, partially unsaturated or
aromatic ring system containing 0-1 heteroatoms selected from O, N,
said ring system optionally substituted with a bond to the linking
group or a bond to the surfactant; and [0861] R.sup.12 is
independently C.sub.1-6 alkyl; [0862] R.sup.27 is .dbd.O, C1-4
alkyl, or phenyl substituted with R.sup.28; [0863] R.sup.28 is a
phenoxy group substituted with 0-2 OCH.sub.3 groups; [0864]
R.sup.29 and R.sup.30 taken together with the nitrogen atom through
which they are attached form a C5-7 atom saturated ring system
substituted with R.sup.31; and [0865] R.sup.31 is a benzyloxy group
substituted with C1-4 alkyl. (57) A diagnostic agent according to
any one of embodiments 55-57 wherein the targeting moiety is a
matrix metalloproteinase inhibitor of the formulae (Ia) or (Ib):
wherein: [0866] R is --OH; [0867] R.sup.2 is C.sub.1-6 alkyl;
[0868] X is C.dbd.O; [0869] R.sup.3 is
[0869] ##STR00063## [0870] R.sup.1 and R.sup.4 are taken together
to form a bridging group of formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.2--; [0871] R.sup.5 is
NH(C1-6alkyl), substituted with a bond to the linking group or a
bond to the surfactant. (58) A diagnostic agent according to any
one of embodiments 55-57 wherein: [0872] R is --OH; [0873] R.sup.9
is C.sub.1 alkyl substituted with a bond to Ln; [0874] R.sup.10 and
R.sup.11 taken together with the nitrogen atom to which they are
attached form a 5 atom saturated ring system, said right system is
substituted with 0-3 R.sup.27; [0875] R.sup.27 is .dbd.O, C1-4
alkyl, or phenyl substituted with R.sup.28; and [0876] R.sup.28 is
a phenoxy group substituted with 0-2 OCH.sub.3 groups. (59) A
diagnostic agent according to any one of embodiments 55-58
wherein
R is --OH;
[0877] R.sup.1 and R.sup.2 taken together with the nitrogen and
carbon atom through which they are attached form a C.sub.5-7 atom
saturated ring system substituted with one or more substituents
selected from the group consisting of: a bond to Ln, a bond to Sf,
and --C(.dbd.O)--NR.sup.29R.sup.30; R.sup.29 and R.sup.30 taken
together with the nitrogen atom through which they are attached
form a C5-7 atom saturated ring system substituted with R.sup.31;
and R.sup.31 is a benzyloxy group substituted with C1-4 alkyl. (60)
A diagnostic agent according to any one of embodiments 48-59
wherein the linking group is of the formula:
((W.sup.1).sub.h-(CR.sup.13R.sup.14).sub.g).sub.x--(Z).sub.k--((CR.sup.1-
3aR.sup.14a).sub.g'-(W.sup.2).sub.h').sub.x'; [0878] W.sup.1 and
W.sup.2 are independently selected at each occurrence from the
group: O, S, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2, SO.sub.2NH,
--(OCH.sub.2CH.sub.2).sub.76-84, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0879] as is
independently at each occurrence an amino acid; [0880] Z is
selected from the group: aryl substituted with 0-3 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.16; [0881] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, PO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-3 R.sup.16, aryl substituted with 0-3 R.sup.16,
benzyl substituted with 0-3 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-3 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to the surfactant; [0882] R.sup.16 is independently
selected at each occurrence from the group: a bond to the
surfactant, COOR.sup.17, C(.dbd.O)NHR.sup.17, NHC(.dbd.O)R.sup.17,
OH, NHR.sup.17, SO.sub.3H, PO.sub.3H, --OPO.sub.3H.sub.2,
--OSO.sub.3H, aryl substituted with 0-3 R.sup.17, C.sub.1-5 alkyl
substituted with 0-1 R.sup.18, C.sub.1-5 alkoxy substituted with
0-1 R.sup.18, and a 5-10 membered heterocyclic ring system
containing 1-4 heteroatoms independently selected from N, S, and O
and substituted with 0-3 R.sup.17; [0883] R.sup.17 is independently
selected at each occurrence from the group: H, alkyl substituted
with 0-1 R.sup.18, aryl substituted with 0-1 R.sup.18, a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-1
R.sup.18, C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.18,
polyalkylene glycol substituted with 0-1 R.sup.18, carbohydrate
substituted with 0-1 R.sup.18, cyclodextrin substituted with 0-1
R.sup.18, amino acid substituted with 0-1 R.sup.18,
polycarboxyalkyl substituted with 0-1 R.sup.18, polyazaalkyl
substituted with 0-1 R.sup.18, peptide substituted with 0-1
R.sup.18, wherein the peptide is comprised of 2-10 amino acids,
3,6-O-disulfo-B-D-galactopyranosyl, bis(phosphonomethyl)glycine,
and a bond to the surfactant; [0884] R.sup.18 is a bond to the
surfactant; [0885] k is selected from 0, 1, and 2; [0886] h is
selected from 0, 1, and 2; [0887] h' is selected from 0, 1, and 2;
[0888] g is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0889] g' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0890] s is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0891] s' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0892] s'' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0893] t is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0894] t' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[0895] x is selected from 0, 1, 2, 3, 4, and 5; and [0896] x' is
selected from 0, 1, 2, 3, 4, and 5. (61) A diagnostic agent
according to any one of embodiments 48-60 wherein [0897] W.sup.1
and W.sup.2 are independently selected at each occurrence from the
group: O, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2,
--(CH.sub.2CH.sub.2O).sub.76-84--, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [0898] aa is
independently at each occurrence an amino acid; [0899] Z is
selected from the group: aryl substituted with 0-1 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-1
R.sup.16; [0900] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-1 R.sup.16, aryl substituted with 0-1 R.sup.16,
benzyl substituted with 0-1 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-1 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to the surfactant; [0901] k is 0 or 1; [0902] s is
selected from 0, 1, 2, 3, 4, and 5; [0903] s' is selected from 0,
1, 2, 3, 4, and 5; [0904] s'' is selected from 0, 1, 2, 3, 4, and
5; and [0905] t is selected from 0, 1, 2, 3, 4, and 5. (62) A
diagnostic agent according to embodiment 60 wherein: [0906] W.sup.1
is C(.dbd.O)NR.sup.15; [0907] h is 1; [0908] g is 3; [0909]
R.sup.13 and R.sup.14 are independently H; [0910] x is 1; [0911] k
is 0; [0912] g' is 0; [0913] h' is 1; [0914] W.sup.2 is NH; and
[0915] x' is 1. (63) A diagnostic agent according to embodiment 60
[0916] x is 0; [0917] k is 1; [0918] Z is aryl substituted with 0-3
R.sup.16; [0919] g' is 1; [0920] W.sup.2 is NH; [0921] R.sup.13a
and R.sup.14a are independently H; [0922] h' is 1; and [0923] x' is
1. (64) A diagnostic agent according to embodiment 60 [0924]
W.sup.1 is C(.dbd.O)NR.sup.15; [0925] h is 1; [0926] g is 2; [0927]
R.sup.13 and R.sup.14 are independently H; [0928] x is 1; [0929] k
is 0; [0930] g' is 1; [0931] R.sup.13a and R.sup.14a are
independently H; or C1-5 alkyl substituted [0932] with 0-3
R.sup.16; [0933] R.sup.16 is SO.sub.3H; [0934] W.sup.2 is
NHC(.dbd.O) or NH; [0935] h' is 1; and [0936] x' is 2. (65) A
diagnostic agent according to embodiment 60 [0937] W.sup.1 is
C(.dbd.O)NH; [0938] h is 1; [0939] g is 3; [0940] R.sup.13 and
R.sup.14 are independently H; [0941] k is 0; [0942] g' is 0; [0943]
x is 1, [0944] W.sup.2 is --NH(C.dbd.O)-- or
--(OCH.sub.2CH.sub.2).sub.76-84--; [0945] h' is 2; and [0946] x' is
1. (66) A diagnostic agent according to embodiment 60 [0947] x is
0; [0948] k is 0; [0949] g' is 3; [0950] h' is 1; [0951] W.sup.2 is
NH; and [0952] x' is 1. (67) A diagnostic agent according to
embodiment 60 [0953] x is 0; [0954] Z is aryl substituted with 0-3
R.sup.16, [0955] k is 1; [0956] g' is 1; [0957] R.sup.13aR.sup.14a
are independently H; [0958] W.sup.2 is NHC(.dbd.O) or
--(OCH2CH2).sub.76-84-; and [0959] x' is 1. (68) A diagnostic agent
according to embodiment 60 [0960] W.sup.1 is C.dbd.O; [0961] g is
2; [0962] R.sup.13 and R.sup.14 are independently H; [0963] k is 0;
[0964] g' is 0; [0965] h' is 1; [0966] W.sup.2 is NH; and [0967] x'
is 1. (69) A diagnostic agent according to embodiment 48 wherein
the linking group is present. (70) A diagnostic agent according to
any one of embodiments 48-69 wherein [0968] S.sub.f is a surfactant
which is a lipid or a compound of the formula:
[0968] ##STR00064## [0969] A.sup.9 is selected from the group: OH
and OR.sup.32; [0970] A.sup.10 is OR.sup.32; [0971] R.sup.32 is
C(O)C.sub.1-20 alkyl; [0972] E.sup.9 is C.sub.1-10 alkylene
substituted with 1-3 R.sup.33; [0973] R.sup.33 is independently
selected at each occurrence from the group: [0974] R.sup.35,
--PO.sub.3H--R.sup.35, .dbd.O, --CO.sub.2R.sup.34,
--C(.dbd.O)R.sup.34, --C(.dbd.O)N(R.sup.34).sub.2,
--CH.sub.2OR.sup.34, --OR.sup.34, --N(R.sup.34).sub.2,
C.sub.1-C.sub.5 alkyl, and C.sub.2-C.sub.4 alkenyl; [0975] R.sup.34
is independently selected at each occurrence from the group:
R.sup.35, H, C.sub.1-C.sub.6 alkyl, phenyl, benzyl, and
trifluoromethyl; [0976] R.sup.35 is a bond to L.sub.n; [0977] and a
pharmaceutically acceptable salt thereof. (71) A diagnostic agent
according to any one of embodiments 48-70 wherein the surfactant is
a lipid or a compound of the formula:
[0977] ##STR00065## [0978] A.sup.9 is OR.sup.32, [0979] A.sup.10 is
OR.sup.32; [0980] R.sup.32 is C(.dbd.O)C.sub.1-15 alkyl, [0981]
E.sup.9 is C.sub.1-4 alkylene substituted with 1-3 R.sup.33; [0982]
R.sup.33 is independently selected at each occurrence from the
group: R.sup.35, --PO.sub.3H--R.sup.35, .dbd.O, --CO.sub.2R.sup.34,
--C(.dbd.O)R.sup.34, --CH.sub.2OR.sup.34, --OR.sup.34, and
C.sub.1-C.sub.5 alkyl; [0983] R.sup.34 is independently selected at
each occurrence from the group: R.sup.35, H, C.sub.1-C.sub.6 alkyl,
phenyl, and benzyl; and [0984] R.sup.35 is a bond to L.sub.n. (72)
A diagnostic agent according to any one of embodiments 48-71,
wherein
##STR00066##
[0984] wherein: [0985] A.sup.1 is a bond to Ln; [0986] E.sup.1 is
C.sub.1 alkyl substituted by R.sup.23; [0987] A.sup.2 is NH; [0988]
E.sup.2 is C.sub.2 alkyl substituted with 0-1R.sup.23; [0989]
A.sup.3 is --O--P(O) (R.sup.21)--O; [0990] E.sup.3 is C.sub.1
alkyl; [0991] A.sup.4 and A.sup.5 are each --O--; [0992] E.sup.4
and E.sup.6 are each independently C.sub.1-16 alkyl substituted
with 0-1R.sup.23; [0993] E.sup.5 is C.sub.1 alkyl; [0994] A.sup.5
is --O--; [0995] R.sup.21 is --OH; and [0996] R.sup.23 is .dbd.O.
(73) A diagnostic agent according to embodiment 48 wherein the
compound is of the formula:
[0996] (Q).sub.d-L.sub.n-S.sub.f
wherein, Q is a compound of Formulae (Ia) or (Ib):
##STR00067##
wherein, [0997] R is independently OH or --CH.sub.2SH; [0998]
R.sup.1 is independently selected at each occurrence from the
group: H, OH, C.sub.1-3 alkyl, C.sub.2-3 alkenyl, C.sub.2-3
alkynyl, and heterocycle-S--CH.sub.2--; [0999] R.sup.2 is
independently C.sub.1-20 alkyl; [1000] X is independently C.dbd.O
or SO.sub.2, provided when X is C.dbd.O, R.sup.3 is
##STR00068##
[1000] and when X is SO.sub.2, R.sup.3 is independently selected
from the group: aryl substituted with 0-2 R.sup.6, and heterocycle
substituted with 0-2 R.sup.6; [1001] R.sup.4 is independently
selected at each occurrence from the group: C.sub.1-6 alkyl,
phenyl, and benzyl; [1002] R.sup.5 is independently at each
occurrence from the group: NH(C.sub.1-6 alkyl), NH-phenyl, and
NH-heterocycle; wherein said alkyl, phenyl and heterocycle groups
are optionally substituted with a bond to L.sub.n; [1003] R.sup.6
is independently aryloxy substituted with 0-3 R.sup.7; [1004]
R.sup.7 is independently halogen or methoxy; [1005] or
alternatively, [1006] R.sup.1 and R.sup.4 may be taken together to
form a bridging group of the formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.2--, optionally substituted
with a bond to L.sub.n; [1007] or alternatively, [1008] R.sup.1 and
R.sup.2 may be taken together to form a bridging group of the
formula --(CH.sub.2).sub.3--NH--, optionally substituted with a
bond to L.sub.n; or [1009] R.sup.1 and R.sup.2 taken together with
the nitrogen and carbon atom through which they are attached form a
C.sub.5-7 atom saturated ring system substituted with one or more
substituents selected from the group consisting of: a bond to Ln, a
bond to Sf, and --C(.dbd.O)--NR.sup.29R.sup.30; [1010] R.sup.8 is
independently at each occurrence OH or phenyl, optionally
substituted with a bond to L.sub.n, provided that when R.sup.8 is
phenyl, R.sup.10 is --C(.dbd.O)--CR.sup.12--NH--CH(CH.sub.3)--COOH;
[1011] R.sup.9 and R.sup.9' are independently H, C.sub.1-6 alkyl
optionally substituted with a bond to L.sub.n, or are taken
together with the carbon atom to which they are attached to form a
5-7 atom saturated, partially unsaturated or aromatic ring system
containing 0-3 heteroatoms selected from O, N, SO.sub.2 and S, said
ring system substituted with R.sup.6 and optionally substituted
with a bond to L.sub.n; [1012] R.sup.10 and R.sup.11 are
independently H, or C.sub.1-6 alkyl optionally substituted with a
bond to L.sub.n, or are taken together with the nitrogen atom to
which they are attached to form a 5-7 atom saturated, partially
unsaturated or aromatic ring system containing 0-3 heteroatoms
selected from O, N, SO.sub.2 and S, said ring system optionally
substituted with 0-3 R.sup.27 or a bond to L.sub.n; [1013] or
alternatively, [1014] R.sup.9 and R.sup.10 are taken together with
the carbon atom to which they are attached to form a 5-7 atom
saturated, partially unsaturated or aromatic ring system containing
0-3 heteroatoms selected from O, N, SO.sub.3 and S, said ring
system optionally substituted with a bond to L.sub.n; [1015]
R.sup.12 is independently C.sub.1-20 alkyl; [1016] d is selected
from 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10; [1017] L.sub.n is a linking
group having the formula:
[1017]
((W.sup.1).sub.h-(CR.sup.13R.sup.14).sub.g).sub.x--(Z).sub.k--((C-
R.sup.13aR.sup.14a).sub.g'-(W.sup.2).sub.h').sub.x'; [1018] W.sup.1
and W.sup.2 are independently selected at each occurrence from the
group: O, S, NH, NHC(.dbd.O), C(.dbd.O)NH, NR.sup.15C(.dbd.O),
C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O, OC(.dbd.O),
NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2, SO.sub.2NH,
--(OCH.sub.2CH.sub.2).sub.76-84, (OCH.sub.2CH.sub.2).sub.s,
(CH.sub.2CH.sub.2O).sub.s', (OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [1019] aa is
independently at each occurrence an amino acid; [1020] Z is
selected from the group: aryl substituted with 0-3 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-3 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.16; [1021] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, PO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-3 R.sup.16, aryl substituted with 0-3 R.sup.16,
benzyl substituted with 0-3 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-3 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, R.sup.17,
and a bond to Sf; [1022] R.sup.16 is independently selected at each
occurrence from the group: a bond to Sf, COOR.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)R.sup.17, OH, NHR.sup.17,
SO.sub.3H, PO.sub.3H, --OPO.sub.3H.sub.2, --OSO.sub.3H, aryl
substituted with 0-3 R.sup.17, C.sub.1-5 alkyl substituted with 0-1
R.sup.18, C.sub.1-5 alkoxy substituted with 0-1 R.sup.18, and a
5-10 membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-3
R.sup.17; [1023] R.sup.17 is independently selected at each
occurrence from the group: H, alkyl substituted with 0-1 R.sup.18,
aryl substituted with 0-1 R.sup.18, a 5-10 membered heterocyclic
ring system containing 1-4 heteroatoms independently selected from
N, S, and O and substituted with 0-1 R.sup.18, C.sub.3-10
cycloalkyl substituted with 0-1 R.sup.18, polyalkylene glycol
substituted with 0-1 R.sup.18, carbohydrate substituted with 0-1
R.sup.18, cyclodextrin substituted with 0-1 R.sup.18, amino acid
substituted with 0-1 R.sup.18, polycarboxyalkyl substituted with
0-1 R.sup.18, polyazaalkyl substituted with 0-1 R.sup.18, peptide
substituted with 0-1 R.sup.18, wherein the peptide is comprised of
2-10 amino acids, 3,6-O-disulfo-B-D-galactopyranosyl,
bis(phosphonomethyl)glycine, and a bond to Sf; [1024] R.sup.18 is a
bond to Sf; [1025] k is selected from 0, 1, and 2; [1026] h is
selected from 0, 1, and 2; [1027] h' is selected from 0, 1, and 2;
[1028] g is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[1029] g' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[1030] s is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[1031] s' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[1032] s'' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[1033] t is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[1034] t' is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10;
[1035] x is selected from 0, 1, 2, 3, 4, and 5; [1036] x' is
selected from 0, 1, 2, 3, 4, and 5; [1037] S.sub.f is a surfactant
which is a lipid or a compound of the formula:
[1037] ##STR00069## [1038] A.sup.9 is selected from the group: OH
and OR.sup.32; [1039] A.sup.10 is OR.sup.32; [1040] R.sup.32 is
C(.dbd.O)C.sub.1-20 alkyl; [1041] E.sup.9 is C.sub.1-10 alkylene
substituted with 1-3 R.sup.33: [1042] R.sup.33 is independently
selected at each occurrence from the group: R.sup.35,
--PO.sub.3H--R.sup.35, .dbd.O, --CO.sub.2R.sup.34,
--C(.dbd.O)R.sup.34, --C(.dbd.O)N(R.sup.34).sub.2,
--CH.sub.2OR.sup.34, --OR.sup.34, --N(R.sup.34).sub.2,
C.sub.1-C.sub.5 alkyl, and C.sub.2-C.sub.4 alkenyl; [1043] R.sup.34
is independently selected at each occurrence from the group:
R.sup.35, H, C.sub.1-C.sub.6 alkyl, phenyl, benzyl, and
trifluoromethyl; [1044] R.sup.35 is a bond to L.sub.n; or [1045] Sf
is of the formula:
##STR00070##
[1045] wherein: [1046] A.sup.1 is a bond to Ln; [1047] E.sup.1 is
C.sub.1 alkyl substituted by R.sup.23; [1048] A.sup.2 is NH; [1049]
E.sup.2 is C.sub.2 alkyl substituted with 0-1R.sup.23; [1050]
A.sup.3 is --O--P(O) (R.sup.21)--O; [1051] E.sup.3 is C.sub.1
alkyl; [1052] A.sup.4 and A.sup.5 are each --O--; [1053] E.sup.4
and E.sup.6 are each independently C.sub.1-16 alkyl substituted
with 0-1R.sup.3; [1054] E.sup.5 is C.sub.1 alkyl; [1055] A.sup.5 is
--O--; [1056] R.sup.21 is --OH; and [1057] R.sup.23 is .dbd.O; or
[1058] a pharmaceutically acceptable salt thereof. (74) A
diagnostic agent according to embodiment 73, wherein: [1059] R is
--OH; [1060] R.sup.2 is C1-6 alkyl; [1061] X is C.dbd.O; [1062]
R.sup.3 is
[1062] ##STR00071## [1063] R.sup.1 and R.sup.4 are taken together
to form a bridging group of formula
--(CH.sub.2).sub.3--O-phenyl-CH.sub.2--; [1064] R.sup.5 is
NH(C1-6alkyl), substituted with a bond to the linking group or a
bond to the surfactant. (75) A diagnostic agent according to any
one of embodiments 73-74, wherein: [1065] R is --OH; [1066] R.sup.9
is C.sub.1 alkyl substituted with a bond to Ln;
[1067] R.sup.10 and R.sup.11 taken together with the nitrogen atom
to which they are attached form a 5 atom saturated ring system,
said right system is substituted with 0-3 R.sup.27; [1068] R.sup.27
is .dbd.O, C.sub.1-4 alkyl, or phenyl substituted with R.sup.28;
and [1069] R.sup.28 is a phenoxy group substituted with 0-2
OCH.sub.3 groups; [1070] S.sub.f is a surfactant which is a lipid
or a compound of the formula:
[1070] ##STR00072## [1071] A.sup.9 is OR.sup.32; [1072] A.sup.10 is
OR.sup.32; [1073] R.sup.32 is C(.dbd.O)C.sub.1-15 alkyl; [1074]
E.sup.9 is C.sub.1-4 alkylene substituted with 1-3 R.sup.33; [1075]
R.sup.33 is independently selected at each occurrence from the
group: R.sup.35, --PO.sub.3H--R.sup.35, .dbd.O, --CO.sub.2R.sup.34,
--C(.dbd.O)R.sup.34, --CH.sub.2OR.sup.34, --OR.sup.34, and
C.sub.1-C.sub.5 alkyl; [1076] R.sup.34 is independently selected at
each occurrence from the group: R.sup.35, H, C.sub.1-C.sub.6 alkyl,
phenyl, and benzyl; and R.sup.35 is a bond to L.sub.n. (76) A
diagnostic agent according to any one of embodiments 73-75,
wherein: [1077] R is --OH; [1078] R.sup.9 is C.sub.1 alkyl
substituted with a bond to Ln; [1079] R.sup.10 and R.sup.11 taken
together with the nitrogen atom to which they are attached form a 5
atom saturated ring system, said right system is substituted with
0-3 R.sup.27; [1080] R.sup.27 is .dbd.O, C.sub.1-4 alkyl, or phenyl
substituted with R.sup.28; and [1081] R.sup.28 is a phenoxy group
substituted with 0-2 OCH.sub.3 groups; [1082] S.sub.f is a
surfactant which is a lipid or a compound of the of the
formula:
##STR00073##
[1082] wherein: [1083] A.sup.1 is a bond to Ln; [1084] E.sup.1 is
C.sub.1 alkyl substituted by R.sup.23; [1085] A.sup.2 is NH; [1086]
E.sup.2 is C.sub.2 alkyl substituted with 0-1R.sup.23; [1087]
A.sup.3 is --O--P(O)(R.sup.21)--O; [1088] E.sup.3 is C.sub.1 alkyl;
[1089] A.sup.4 and A.sup.5 are each --O--; [1090] E.sup.4 and
E.sup.6 are each independently C.sub.1-16 alkyl substituted with
0-1R.sup.23; [1091] E.sup.5 is C.sub.1 alkyl; [1092] A.sup.5 is
--O--; [1093] R.sup.21 is --OH; and [1094] R.sup.23 is .dbd.O. (77)
A diagnostic agent according to any one of embodiments 73-76,
wherein: wherein [1095] R is --OH; [1096] R.sup.1 and R.sup.2 taken
together with the nitrogen and carbon atom through which they are
attached form a C.sub.5-7 atom saturated ring system substituted
with one or more substituents selected from the group consisting
of: a bond to Ln, a bond to Sf, and --C(.dbd.O)--NR.sup.29R.sup.30;
[1097] R.sup.29 and R.sup.30 taken together with the nitrogen atom
through which they are attached form a C5-7 atom saturated ring
system substituted with R.sup.31; and [1098] R.sup.31 is a
benzyloxy group substituted with C.sub.1-4 alkyl. [1099] d is
selected from 1, 2, 3, 4, and 5; [1100] W is independently selected
at each occurrence from the group: O, NH, NHC(.dbd.O), C(.dbd.O)NH,
NR.sup.15C(.dbd.O), C(.dbd.O)NR.sup.15, C(.dbd.O), C(.dbd.O)O,
OC(.dbd.O), NHC(.dbd.S)NH, NHC(.dbd.O)NH, SO.sub.2,
(OCH.sub.2CH.sub.2).sub.s, (CH.sub.2CH.sub.2O).sub.s',
(OCH.sub.2CH.sub.2CH.sub.2).sub.s'',
(CH.sub.2CH.sub.2CH.sub.2O).sub.t, and (aa).sub.t'; [1101] aa is
independently at each occurrence an amino acid; [1102] Z is
selected from the group: aryl substituted with 0-1 R.sup.16,
C.sub.3-10 cycloalkyl substituted with 0-1 R.sup.16, and a 5-10
membered heterocyclic ring system containing 1-4 heteroatoms
independently selected from N, S, and O and substituted with 0-1
R.sup.16; [1103] R.sup.13, R.sup.13a, R.sup.14, R.sup.14a, and
R.sup.15 are independently selected at each occurrence from the
group: H, .dbd.O, COOH, SO.sub.3H, C.sub.1-C.sub.5 alkyl
substituted with 0-1 R.sup.16, aryl substituted with 0-1 R.sup.16,
benzyl substituted with 0-1 R.sup.16, and C.sub.1-C.sub.5 alkoxy
substituted with 0-1 R.sup.16, NHC(.dbd.O)R.sup.17,
C(.dbd.O)NHR.sup.17, NHC(.dbd.O)NHR.sup.17, NHR.sup.17, NHR.sup.17,
R.sup.17, and a bond to Sf; [1104] k is 0 or 1; [1105] s is
selected from 0, 1, 2, 3, 4, and 5; [1106] s' is selected from 0,
1, 2, 3, 4, and 5; [1107] s'' is selected from 0, 1, 2, 3, 4, and
5; and [1108] t is selected from 0, 1, 2, 3, 4, and 5. (78) A
diagnostic agent according to according to any one of embodiments
73-77, wherein: [1109] W.sup.1 is C(.dbd.O)NR.sup.15; [1110] h is
1; [1111] g is 3; [1112] R.sup.13 and R.sup.14 are independently H;
[1113] x is 1; [1114] k is 0; [1115] g' is 0; [1116] h' is 1;
[1117] W.sup.2 is NH; and [1118] x' is 1. (79) A diagnostic agent
according to embodiment 73, wherein: [1119] x is 0; [1120] k is 1;
[1121] Z is aryl substituted with 0-3 R.sup.16; [1122] g' is 1;
[1123] W.sup.2 is NH; [1124] R.sup.13a and R.sup.14a are
independently H; [1125] h' is 1; and [1126] x' is 1. (80) A
diagnostic agent according to Embodiment 73, wherein: [1127]
W.sup.1 is C(.dbd.O)NR.sup.15, [1128] h is 1; [1129] g is 2; [1130]
R.sup.13 and R.sup.14 are independently H; [1131] x is 1; [1132] k
is 0; [1133] g' is 1; [1134] R.sup.13a and R.sup.14a are
independently H; or C.sub.1-5 alkyl substituted [1135] with 0-3
R.sup.16; [1136] R.sup.16 is SO.sub.3H; [1137] W.sup.2 is
NHC(.dbd.O) or NH; [1138] h' is 1; and [1139] x' is 2. (81) A
diagnostic agent according to Embodiment 73, wherein: [1140]
W.sup.1 is C(.dbd.O)NH; [1141] h is 1; [1142] g is 3; [1143]
R.sup.13 and R.sup.14 are independently H; [1144] k is 0; [1145] g'
is 0; [1146] x is 1; [1147] W.sup.2 is --NH(C.dbd.O)-- or
--(OCH.sub.2CH.sub.2).sub.76-84--; [1148] h' is 2; and [1149] x' is
1. (82) A diagnostic agent according to Embodiment 73, wherein:
[1150] x is 0; [1151] k is 0; [1152] g' is 3; [1153] h' is 1;
[1154] W.sup.2 is NH; and [1155] x' is 1. (83) A diagnostic agent
according to Embodiment 73, wherein: [1156] x is 0; [1157] Z is
aryl substituted with 0-3 R.sup.16; [1158] k is 1; [1159] g' is 1;
[1160] R.sup.13aR.sup.14a are independently H; [1161] W.sup.2 is
NHC(.dbd.O) or --(OCH2CH2).sub.76-84-; and [1162] x' is 1. (84) A
diagnostic agent according to Embodiment 73, wherein: [1163]
W.sup.1 is C.dbd.O; [1164] g is 2; [1165] R.sup.13 and R.sup.14 are
independently H; [1166] k is 0; [1167] g' is 0; [1168] h' is 1;
[1169] W.sup.2 is NH; and [1170] x' is 1. (85) A diagnostic agent
according to Embodiment 1, wherein the compound is selected from
the group consisting of:
##STR00074##
[1170] (86) A diagnostic agent according to embodiment 48, wherein:
wherein the echogenic gas is a perfluorocarbon gas or sulfur
hexafluoride. (87) A diagnostic agent according to embodiment 86
wherein said perfluorocarbon is selected from the group consisting
of perfluoromethane, perfluoroethane, perfluoropropane,
perfluorobutane, perfluorocyclobutane, perfluoropentane, and
perfluorohexane. (88) A diagnostic composition comprising a
compound according to embodiment 48 or a pharmaceutically
acceptable salt thereof, and a pharmaceutically acceptable carrier.
(89) A diagnostic composition comprising a compound according to
embodiment 48 or a pharmaceutically acceptable salt form thereof,
an echogenic gas and a pharmaceutically acceptable carrier. (90) A
diagnostic composition comprising a compound according to
embodiment 48 further comprising:
1,2-dipalmitoyl-an-glycero-3-phosphotidic acid,
1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine, and
N-(methoxypolyethylene glycol 5000
carbamoyl)-1,2-dipalmitoyl-sn-glycero-3-phosphatidylethanolamine.
c. Third Non-Limiting Set of Embodiments of Imaging Agents or
Precursors Thereof
[1171] Matrix metalloproteinase (MMP) activity and extracellular
matrix degradation is dependent on the comparative balance between
MMPs and TIMPs. Elevated TIMP activity suppresses angiogenesis via
inhibition of endothelial cell migration. TIMPs and synthetic small
molecules or matrix metalloproteinase inhibitors have therapeutic
potential for diseases involving elevated levels of MMP activity
(Whittaker, M. et al, Chem. Rev., 1999, 99, 2735-2776; Babine, R.
E. et al, Chem. Rev., 1997, 97, 1359; De, B. et al, Ann. N.Y. Acad.
Sci., 1999, 878, 40-60; Summers, J. B. et al, Annual Reports in
Med. Chem., 1998, 33, 131).
[1172] A functional group, such as --CONH--OH, --COOH, or --SH, is
necessary for a molecule to be an effective inhibitor of MMPs. This
functional group is involved in the chelation of the active site
zinc ion, and is commonly referred to as the zinc binding group or
ZBG. The hydroxamate, for example, is a bidentate ligand for
zinc.
[1173] In some embodiments, a compound comprises the formula,
(Q).sub.d-L.sub.n-(C.sub.h--X),
(Q).sub.d-L.sub.n-(C.sub.n--X.sup.1).sub.d',
(Q).sub.d-L.sub.n-(C.sub.h--X.sup.2).sub.d'', or
(Q).sub.d-L.sub.n-(C.sub.h--X.sup.3), wherein Q represents a
compound that inhibits a matrix metalloproteinase, d is 1-10,
d'=1-100, d'' is 1-100, Ln represents an optional linking group,
C.sub.h represents a metal chelator or bonding moiety, X represents
a radioisotope, X.sup.1 represents paramagnetic metal ion, X.sup.2
represents a paramagnetic metal ion or heavy atom containing
insoluble solid particle, and X.sup.3 represents a surfactant
microsphere of an echogenic gas.
[1174] One class of compounds that that inhibits a matrix
metalloproteinase (e.g., Q) are succinyl hydroxamates. A generic
structure of succinyl hydroxamate is shown below (1-1).
##STR00075##
[1175] The ethylene spacer between the ZBG (--CONH--OH) and the
succinyl amide is essential for potent activity. Substitution at
P.sub.1 tends to confer broad-spectrum activity on the MMPIs.
Substituents at this position, in general, tend to point away from
the enzyme. Moieties capable of hydrogen bonding and lipophilic
substituents at the P.sub.1 position .alpha. to the hydroxamate
(Johnson, W. H. et al, J. Enz. Inhib., 1987, 2, 1) tend to enhance
activity (1-2). Incorporation of a hydroxyl group (Beckett, P. R.,
et al, Drug Discovery Today, 1996, 1, 16) at that position improves
oral activity in some case (1-3).
##STR00076##
[1176] Substituents at the P.sub.1' position on the succinyl
hydroxamates tend to impart selectivity to the MMPIs. The S.sub.1'
pocket is deep for MMP-2, MMP-3, MMP-8 and MMP-9 and occluded
(short) for MMP-1 and MMP-7. A long alkyl substituent at the
P.sub.1' position, for example, imparts selectivity (Miller, A. et
al, Bioorg. Med. Chem. Lett., 1997, 7, 193) for MMP-2 over MMP-1
and MMP-3 (1-4 and 1-5).
##STR00077##
[1177] Substituents at the P.sub.2' position also point away from
the enzyme. The P.sub.1 and the P.sub.2' positions can be linked
(Xue, C-B. et al, J. Med. Chem., 1998, 41, 1745; Steinman, D. H. et
al, Bioorg. Med. Chem. Lett., 1998, 8, 2087) to form a macrocycle
(1-6). Compounds such as (1-6) also exhibit nanomolar activity.
##STR00078##
[1178] The nature of the macrocycle also imparts some selective
inhibition among the MMPs. The P.sub.2' and the P.sub.3' positions
may be cyclized to form lactams. The size of the lactam governs the
selectivity.
[1179] The P.sub.3' position is a relatively open area in the
succinyl hydroxamates, and a wide range of substitutents (for
example (1-7)) may be introduced (Sheppard, G. S. et al, Bioorg.
Med. Chem. Lett., 1998, 8, 3251) at this position. This position
also offers the flexibility of attaching the optional linker,
L.sub.n, the chelator(s0, C.sub.h, for the imageable moieties X and
X.sup.1, and the imageable moieties, X.sup.2 and X.sup.3.
##STR00079##
[1180] Other succinyl hydroxamates with modified P.sub.2' and
P.sub.3' positions, such as (1-8) also have shown potent inhibition
of MMPs. Those compounds and syntheses of them are further
described in the following patent applications which are hereby
incorporated by reference into this patent application: U.S. patent
application Ser. No. 08/743,439, 60/127,594, and 60/127,635 and
U.S. Pat. Nos. 6,057,336, 6,576,664, 6,455,522, 6,429,213,
6,365,587, 6,268,379, 6,495,548, 6,689,771, and 6,376,665
##STR00080##
[1181] Another class of compounds of that inhibits a matrix
metalloproteinase (e.g., Q) are sulfonamide hydroxamates, such as
(1-9) and (1-10). Modification of the isopropyl substituent in
(1-10) results in deep pocket MMP selectivity, for example MMP-2 vs
MMP-1 (Santos, O. et al., J. Clin. Exp. Metastasis, 1997, 15, 499;
MacPherson, L. J. et al, J. Med. Chem., 1997, 40, 2525).
##STR00081##
[1182] Additional examples of inhibitors, Q, include the
derivatized alanine hydroxamates, such as compounds (1-11) and
(1-12), which show selectivity for MMP-2 and MMP-9 over the other
MMPs. The P.sub.1 position is available for limited modification as
described in the patents and applications incorporated by reference
above. Those compounds and syntheses of them are further described
in the following patent applications which are hereby incorporated
by reference into this patent application: U.S. patent application
Ser. No. 08/743,439, 60/127,594, and 60/127,635 and U.S. Pat. Nos.
6,057,336, 6,576,664, 6,455,522, 6,429,213, 6,365,587, 6,268,379,
6,495,548, 6,689,771, and 6,376,665
##STR00082##
[1183] Other compounds with selectivity for MMP-2 and MMP-9 over
MMP-1 include (1-13). In this example the alpha position has a
quaternary carbon and the molecule does not contain any stereo
centers (Lovejoy, B. et al., Nature Struct. Biol., 1999, 6,
217).
##STR00083##
[1184] In the non-hydroxamate series, a number of compounds have
been reported with a variety of structures. Use of carboxylic acid
as the ZBG has also received attention. In the case of compound
(1-14), significant selectivity for MMP-2 (vs MMP-1) was observed
when X=butyl vs X=H (Sahoo, S. P. et al, Bioorg. Med. Chem. Lett.,
1995, 5, 2441).
##STR00084##
[1185] Although thiols are monodentate ZBGs, some succinyl thiols
such as (1-15) have exhibited good activity (Levin, J. I. et al,
Bioorg. Med. Chem. Lett., 1998, 8, 1163). The P.sub.3' position may
be utilized to conjugate a variety of linkers and chelators (as
described above) for the preparation of diagnostic agents. For
example, the P.sub.3' position may be utilized to attach the
optional linker, L.sub.n, the chelator(s), C.sub.h, for the
imageable moieties X and X.sup.1, and the imageable moieties,
X.sup.2 and X.sup.3
##STR00085##
[1186] In some embodiments, the pharmaceuticals are comprised of
inhibitors, Q, which exhibit selectivity for MMP-1, MMP-2, MMP-3,
MMP-9, or MMP-14 alone or in combination over the other MMPs.
Examples of moieties, Q, include compounds 1-4, 1-5, 1-6, 1-8, 1-9,
1-10, 1-11, 1-12, and 1-13.
[1187] In some embodiments, the inhibitors, Q, is selected to
exhibit selectivity for MMP-2, MMP-9, or MMP-14 alone or in
combination over the other MMPs. Examples of the such moieties, Q,
include compounds 1-6, 1-8, 1-11, and 1-12.
[1188] Such pharmaceuticals can be synthesized by several
approaches. One approach involves the synthesis of the targeting
MMP inhibiting moiety, Q, and direct attachment of one or more
moieties, Q, to one or more metal chelators or bonding moieties,
C.sub.h, or to a paramagnetic metal ion or heavy atom containing
solid particle, or to an echogenic gas microbubble. Another
approach involves the attachment of one or more moieties, Q, to the
linking group, L.sub.n, which is then attached to one or more metal
chelators or bonding moieties, C.sub.h, or to a paramagnetic metal
ion or heavy atom containing solid particle, or to an echogenic gas
microbubble. Another approach, useful in the synthesis of
pharmaceuticals wherein d is 1, involves the synthesis of the
moiety, Q-L.sub.n, together, by incorporating residue bearing
L.sub.n into the synthesis of the MMP inhibitor, Q. The resulting
moiety, Q-L.sub.n, is then attached to one or more metal chelators
or bonding moieties, C.sub.h, or to a paramagnetic metal ion or
heavy atom containing solid particle, or to an echogenic gas
microbubble. Another approach involves the synthesis of an
inhibitor, Q, bearing a fragment of the linking group, L.sub.n, one
or more of which are then attached to the remainder of the linking
group and then to one or more metal chelators or bonding moieties,
C.sub.h, or to a paramagnetic metal ion or heavy atom containing
solid particle, or to an echogenic gas microbubble.
[1189] The MMP inhibiting moieties, Q, optionally bearing a linking
group, L.sub.n, or a fragment of the linking group, can be
synthesized using standard synthetic methods known to those skilled
in the art. Methods include but are not limited to those methods
described below.
[1190] Generally, peptides, polypeptides and peptidomimetics are
elongated by deprotecting the alpha-amine of the C-terminal residue
and coupling the next suitably protected amino acid through a
peptide linkage using the methods described. This deprotection and
coupling procedure is repeated until the desired sequence is
obtained. This coupling can be performed with the constituent amino
acids in a stepwise fashion, or condensation of fragments (two to
several amino acids), or combination of both processes, or by solid
phase peptide synthesis according to the method originally
described by Merrifield, J. Am. Chem. Soc., 85, 2149-2154 (1963),
the disclosure of which is hereby incorporated by reference.
d. Fourth Non-Limiting Set of Embodiments of Imaging Agents or
Precursors Thereof
[1191] In some embodiments, an imaging agent or imaging agent
precursor is selected from the group consisting of:
##STR00086## ##STR00087## ##STR00088## ##STR00089##
Subjects
[1192] As used herein, "subject" includes, but is not limited to,
vertebrates, more specifically a mammal (e.g., a human, horse, pig,
rabbit, dog, sheep, goat, non-human primate, cow, cat, guinea pig
or rodent), a fish, a bird or a reptile or an amphibian. In some
embodiments, the subject is a human subject. As used herein,
"patient" refers to a subject afflicted with a disease or disorder.
The term "patient" includes human and veterinary subjects.
[1193] In some instances, the subject is one that has not
experienced a cardiovascular insult such as a myocardial
infarction.
[1194] In some instances, the subject is one that has experienced a
myocardial infarction. In some cases, the imaging methods may be
performed within hours, days, weeks, or months after the myocardial
infarction. In some instances, the imaging methods are performed
repeatedly after a myocardial infarction including for example
weekly, monthly, biannually, annually, etc. The imaging methods may
be performed at different frequencies after a myocardial
infarction. As an example, immediately after a myocardial
infarction, the methods may be performed on a weekly or monthly
basis for a period of time (e.g., 6 months or a year), and
thereafter may be performed at a less regular interval (e.g., every
6 months, or every year) for a period of time or indefinitely.
[1195] In some instances, the subject is one having atherosclerosis
(e.g., having symptoms of atherosclerosis). In some instances, the
subject is one that does not have atherosclerosis (e.g., does not
have symptoms of atherosclerosis).
Increased Risk Vs. Normal Population
[1196] The invention contemplates, in part, detecting presence and
in some instances amount of an administered imaging agent (i.e., an
MMP inhibitor linked to an imaging moiety) and comparing this to a
control in order to determine an increased risk of developing AF or
other indication (e.g., CAVD). The invention intends to determine
an "above-normal" or "above-average" or "increased" risk of
developing AF or other indication. An above-normal or above-average
risk or increased risk is a risk that is greater than the risk of a
normal subject or a population of normal subjects or a randomly
selected population for developing AF or other indication. In some
instances, an above-normal or above-average risk or increased risk
is indicated by any level of MMP that is greater than the MMP level
of a control. In some instances, the increased risk is further
quantified by measuring the MMP level, wherein lower MMP levels
indicate a lower "increased" risk and higher MMP levels indicate a
higher "increased" risk, provided that even the lower MMP levels
are still above normal or control levels.
[1197] The control level may be an MMP level determined using the
same imaging agent in a normal subject (i.e., a subject that is
known not to have AF), or it may be the average MMP level in a
population of normal subjects, or it may be the average MMP level
in a random sampling of the population at large. The control level
may be one that is determined prior to the analysis of the subject
rather than one that is determined in real time. The control level
may therefore be a level that is obtained and established on a
periodic basis (e.g., every 6 months, every year, etc.).
[1198] It will be understood that all subjects may have some risk
of developing AF or other indication (e.g., CAVD). This risk may be
referred to herein as the "normal risk." The normal risk may be
established on an individual subject basis or on a population
basis. For example, it may be determined as the risk of a "normal"
subject developing AF (i.e., a subject that is not known to have
AF), or the average risk in a normal population of developing AF,
or the average risk in a randomly selected subpopulation from the
population at large.
[1199] In still other embodiments, the invention further
contemplates determining an increased risk of developing AF based
on clinical trial results. As an example, a clinical trial may be
performed that assesses MMP profile in a number of subjects and
then follows those subjects over time in order to determine the
nature of the profile that correlates with increased risk of AF.
Those trials may be performed on subjects that previously had AF
and that may have been treated with an AF therapy (such as but not
limited to cardioversion), with an outcome of determining a profile
that correlates with subjects that do not have a recurrence of AF
after the trial and/or determining a profile that correlates with
subjects that do have a recurrence of AF after the trial. Such
trials may then be used to set the threshold (or cut point or
control) to which future subjects are compared against.
[1200] Regardless of the control used, increased risk of developing
AF (as a primary event or as a recurrent event) and/or likelihood
of responding to a particular AF therapy (such as but not limited
to cardioversion, ablation, pharmacological rate or rhythm control
therapy, or implantable pacer) may be indicated by any MMP level
that is above a control level, or it may be indicated by an MMP
level that is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,
50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 100%, 200%, 300%,
400%, or 500%, or 10-fold, 20-fold, 50-fold, 100-fold, 200-fold,
300-fold, 400-fold, or 500-fold more than the control level.
Imaging
[1201] The invention contemplates administering, to a subject, an
imaging agent of the invention and then acquiring one or more
images of the subject. The images will typically comprise images of
the subject's heart, in whole or in part. Such images are therefore
referred to herein as heart or cardiac images. Such images may
comprise more than heart tissue and/or they may not comprise the
entire heart tissue. In some instances, such images will comprise
atrial myocardium, including left atrial myocardium.
[1202] The imaging modality will be dictated by the imaging moiety
linked to the MMP inhibitor (and vice versa) as will be understood.
In some embodiments, the imaging modality is single-photon emission
computed tomography (referred to as SPECT or SPET), SPECT/CT, PET,
ultrasound, MRI, and the like.
Myocardial (or Cardiac) Perfusion
[1203] In some instances, the invention further contemplates
determining myocardial perfusion (i.e., blood flow through the
heart) using myocardial perfusion imaging agents. In some
instances, a measure of myocardial perfusion is used together with
a measure of MMP levels. Myocardial perfusion imaging agents
include but are not limited to flurpiridaz F18, Thallium-201, and
Tc-Sestamibi.
[1204] In some cases, methods of the invention may include
determining a parameter of, or the presence or absence of,
myocardial ischemia, rest (R) and/or stress (S) myocardial blood
flows (MBFs), coronary flow reserve (CFR), coronary artery disease
(CAD), left ventricular ejection fraction (LVEF), end-systolic
volume (ESV), end-diastolic volume (EDV), and the like.
Atrial Fibrillation (AF)
[1205] AF is an abnormal heart rhythm (cardiac arrhythmia) which
involves the two small, upper heart chambers (i.e., the atria).
Heart beats in a normal heart begin after electricity generated in
the atria by the sinoatrial node spreads through the heart and
causes contraction of the heart muscle and pumping of blood. In AF,
the regular electrical impulses of the sinoatrial node are replaced
by disorganized, rapid electrical impulses which result in
irregular heartbeats.
[1206] AF is the most common cardiac arrhythmia. An individual may
spontaneously alternate between AF and a normal rhythm (paroxysmal
AF) or may continue with AF as the dominant cardiac rhythm without
reversion to the normal rhythm (chronic AF).
[1207] AF is often asymptomatic, but may result in symptoms of
palpitations, fainting, chest pain, or even heart failure. These
symptoms are especially common when AF results in a heart rate
which is either too fast or too slow. In addition, the erratic
motion of the atria leads to blood stagnation (stasis) which
increases the risk of blood clots that may travel from the heart to
the brain and other areas. Thus, AF is an important risk factor for
stroke, the most feared complication of AF.
[1208] The symptoms of AF may be treated with pharmacological
agents which slow the heart rate. Several such pharmacological
agents as well as electrical cardioversion may be used to convert
AF to a normal heart rhythm. Surgical and catheter-based therapies
may also be used to prevent AF in certain individuals. People with
AF are often given blood thinners such as warfarin to protect them
from strokes.
[1209] The American Heart Association, American College of
Cardiology, and the European Society of Cardiology have proposed
the following classification system based on simplicity and
clinical relevance. "First Detected" refers to any patient newly
diagnosed with AF, as the exact onset and chronicity of the disease
is often uncertain. A patient with 2 or more identified episodes of
AF is said to have "recurrent" AF. This is further classified into
"paroxysmal" and "persistent" based on when the episode terminates
without therapy. AF is said to be "paroxysmal" when it terminates
spontaneously within 7 days, most commonly within 24 hours.
"Persistent" or "chronic" AF is AF established for more than seven
days. Differentiation of paroxysmal from chronic or established AF
is based on the history of recurrent episodes and the duration of
the current episode of AF. "Lone atrial fibrillation" (LAF) is
defined as AF in the absence of clinical or echo cardiographic
findings of cardiopulmonary disease. Patients with LAF who are
under 65 have the best prognosis.
[1210] AF is usually accompanied by symptoms related to either
rapid heart rate or embolization. Rapid and irregular heart rates
may be perceived as palpitations, exercise intolerance, and
occasionally produce angina and congestive symptoms of shortness of
breath or edema. Sometimes the arrhythmia will be identified with
the onset of a stroke or a transient ischemic attack (TIA). It is
not uncommon to identify AF on a routine physical examination or
electrocardiogram (ECG/EKG), as it may be asymptomatic in some
cases. Paroxysmal AF is the episodic occurrence of the arrhythmia
and may be difficult to diagnose. Episodes may occur with sleep or
with exercise, and their episodic nature may require prolonged ECG
monitoring (e.g. a Holter monitor) for diagnosis.
[1211] AF is diagnosed on an electrocardiogram, an investigation
performed routinely whenever irregular heart beat is suspected.
Characteristic findings include absence of P waves, unorganized
electrical activity in their place, and irregularity of R-R
interval due to irregular conduction of impulses to the ventricles.
If paroxysmal AF is suspected, episodes may be documented with the
use of Holter monitoring (continuous ECG recording for 24 hours or
longer).
[1212] Diagnosis of AF sometimes involves analysis of renal
function and electrolytes, as well as thyroid-stimulating hormone
(commonly suppressed in hyperthyroidism and of relevance if
amiodarone is administered for treatment) and a blood count. A
chest X-ray is generally performed. In acute-onset AF associated
with chest pain, cardiac troponins or other markers of damage to
the heart muscle may be ordered. Coagulation studies (INR/aPTT) are
usually performed, as anticoagulant medication may be commenced. A
transesophageal echocardiogram may be indicated to identify any
intracardiac thrombus.
[1213] AF is linked to several cardiac causes, but may occur in
otherwise normal hearts. Known associations include carbon monoxide
poisoning, high blood pressure, mitral stenosis (e.g. due to
rheumatic heart disease or mitral valve prolapse), mitral
regurgitation, heart surgery, coronary artery disease, hypertrophic
cardiomyopathy, excessive alcohol consumption ("binge drinking" or
"holiday heart syndrome"), hyperthyroidism, hyperstimulation of the
vagus nerve, usually by having large meals ("binge eating"), lung
pathology (such as pneumonia, lung cancer, pulmonary embolism,
sarcoidosis), pericarditis, intense emotional turmoil, and
congenital heart disease.
[1214] The main goals of treatment of AF are to prevent temporary
circulatory instability and stroke. Rate control and rhythm control
are principally used to achieve the former, while anticoagulation
may be required to decrease the risk of the latter. AF can cause
disabling and annoying symptoms. Palpitations, angina, lassitude
(weariness), and decreased exercise tolerance are related to rapid
heart rate and inefficient cardiac output caused by AF. Rate
control treatments seek to reduce the heart rate to normal, usually
60 to 100 beats per minute. Rhythm control seeks to restore the
normal heart rhythm, called normal sinus rhythm. Studies suggest
that rhythm control is mainly a concern in newly diagnosed AF,
while rate control is more important in the chronic phase.
[1215] AF with a persistent rapid rate can cause a form of heart
failure called tachycardia induced cardiomyopathy. This can
significantly increase mortality and morbidity. The early treatment
of AF through either rate-control or rhythm control can prevent
this condition and thereby improve mortality and morbidity.
[1216] Rate control methods include beta blockers (e.g.
metoprolol), cardiac glycosides (e.g. digoxin), and calcium channel
blockers (e.g. verapamil). These medications work by slowing the
generation of impulses from the atria and the conduction of those
impulses from the atria to the ventricles.
[1217] In refractory cases where none of the above drugs are
sufficient, a variety of other antiarrhythmic drugs, most commonly
including quinidine, flecamide, propafenone, disopyramide, sotalol,
or amiodarone may be used. Of these, only propafenone, sotalol, and
amiodarone (which possess some beta blocking activity) control the
ventricular rate; the others may maintain sinus rhythm, but may
actually increase the ventricular rate. Many of these drugs are
less frequently used today than in the past. All (with the possible
exception of amiodarone) increase the risk of ventricular
tachycardia, which can be fatal. In symptomatic patients with
normal heart function, however, the small increase in risk is
usually felt to be acceptable. In the presence of heart failure,
the only anti-arrhythmic drugs thought to be safe are amiodarone
and dofetilide.
[1218] In patients with AF where rate control drugs are ineffective
and it is not possible to restore sinus rhythm using cardioversion,
non-pharmacological alternatives are available. For example, to
control rate it is possible to destroy the bundle of cells
connecting the upper and lower chambers of the heart--the
atrioventricular node--which regulates heart rate, and to implant a
pacemaker instead.
[1219] A more complex technique involves ablating groups of cells
near the pulmonary veins where AF is thought to originate, or
creating more extensive lesions in an attempt to prevent AF from
establishing itself.
[1220] Rhythm control methods include electrical and chemical
cardioversion. Electrical cardioversion involves the restoration of
normal heart rhythm through the application of a DC (direct
current) electrical shock. Chemical cardioversion is performed with
drugs, such as amiodarone, propafenone or flecamide. Implantable
pacing devices can also be used for rate management of AF patients
and can be indicated versus traditional cardioversion.
[1221] The anti-arrhythmic medications often used in either
pharmacological cardioversion or in the prevention of relapse to AF
alter the flux of ions in heart tissue, making them less excitable,
setting the stage for spontaneous and durable cardioversion. These
medications are often used in concert with electrical
cardioversion.
[1222] Whichever method of cardioversion is used, approximately 50%
of patients relapse within one year, although the continued daily
use of oral antiarrhythmic drugs may extend this period. The key
risk factor for relapse is duration of AF, although other risk
factors that have been identified include the presence of
structural heart disease, and increasing age.
[1223] Radiofrequency ablation (RFA) uses radiofrequency energy to
destroy abnormal electrical pathways in heart tissue. It is used in
recurrent AF. The energy emitting probe (electrode) is placed into
the heart through a catheter. The practitioner first "maps" an area
of the heart to locate the abnormal electrical activity before the
responsible tissue is eliminated. Ablation is a newer technique and
has shown some promise for cases unresponsive to conventional
treatments. New techniques include the use of cryoablation (tissue
freezing using a coolant which flows through the catheter), and
microwave ablation, where tissue is ablated by the microwave energy
"cooking" the adjacent tissue. The abnormal electrophysiology can
also be modified in a similar way surgically, and this procedure
referred to as the Cox maze procedure, is commonly performed
concomitantly with cardiac surgery. More recently, minimally
invasive surgical variations on the Cox Maze procedure ("minimaze"
procedures) have also been developed.
[1224] The Cox maze procedure is an open-heart surgical procedure
intended to eliminate AF. "Maze" refers to the series of incisions
made in the atria (upper chambers of the heart), which are arranged
in a maze-like pattern. The intention was to eliminate AF by using
incisional scars to block abnormal electrical circuits (atrial
macrorentry) that AF requires. This procedure required an extensive
series of endocardial (from the inside of the heart) incisions
through both atria, a median sternotomy (vertical incision through
the breastbone) and cardiopulmonary bypass (heart-lung machine). A
series of improvements were made, culminating in 1992 in the Cox
maze III procedure, which is now considered to be the "gold
standard" for effective surgical cure of AF. The Cox maze III is
sometimes referred to as the "traditional maze", the "cut and sew
maze", or simply the "maze".
[1225] Minimaze surgery is minimally invasive cardiac surgery
intended to cure AF. Minimaze refers to "mini" versions of the
original maze procedure. These procedures are less invasive than
the Cox maze procedure and do not require a median sternotomy
(vertical incision in the breastbone) or cardiopulmonary bypass
(heart-lung machine). These procedures use microwave,
radiofrequency, or acoustic energy to ablate atrial tissue near the
pulmonary veins.
[1226] In confirmed AF, anticoagulant treatment is a crucial way to
prevent stroke. Treatment of AF patients over age 60, who also have
one or more of: previous strokes (or warning strokes), hypertension
(high blood pressure), diabetes, or congestive heart failure, with
warfarin (also known as Coumadin.RTM. or Marevan.RTM.) results in a
60 to 70 percent reduction in the subsequent risk of stroke.
Patients under age 65 who have any structural heart disease (i.e.
valvular heart disease, ejection fraction <=35%, history of
heart attack) may also benefit from warfarin.
[1227] The use of warfarin is associated with a delayed clinical
effect. It typically takes three to five days to achieve a
demonstrable anticoagulant effect. Hence, if an immediate
anticoagulant effect is required, physicians could use heparin or
other heparinoids such as enoxaparin to provide early
anticoagulation. In practice, urgent anticoagulation is seldom
indicated. Even in the setting of stroke complicating AF, clinical
trial results do not support the routine use of immediate
anticoagulation.
[1228] Patients under age 65 who do not have structural heart
disease (i.e. with LAF) do not require warfarin, and can be treated
with aspirin or clopidogrel. There is evidence that aspirin and
clopidogrel are effective when used together. The new anticoagulant
ximelagatran has been shown to prevent stroke with equal efficacy
as warfarin.
[1229] Determining who should and should not receive
anti-coagulation with anti-coagulant drugs (e.g., warfarin,
ximegalatran, heparin or other heparinoids) is not easy. The CHADS2
score is the best validated method of determining risk of stroke
(and therefore who should be anticoagulated). The UK NICE
guidelines have instead opted for an algorithm approach. The
underlying problem is that if a patient has a yearly risk of stroke
that is less than 2%, then the risks associated with taking
warfarin outweigh the risk of getting a stroke.
MMPs
[1230] Key contributors to ECM synthesis/degradation are the matrix
metalloproteinases (MMPs) and the endogenous tissue inhibitors of
the metalloproteinases (TIMPs) (Visse R, et al. 2003; Matrisian L
M, et al. 1990).
[1231] Matrix metalloproteinases (MMPs) are zinc-dependent
endopeptidases; other family members are adamalysins, serralysins,
and astacins. The MMPs belong to a larger family of proteases known
as the metzincin superfamily.
[1232] The MMPs share a common domain structure. The three common
domains are the pro-peptide, the catalytic domain and the
haemopexin-like C-terminal domain which is linked to the catalytic
domain by a flexible hinge region.
[1233] The MMPs are initially synthesized as inactive zymogens with
a pro-peptide domain that must be removed before the enzyme is
active. The pro-peptide domain is part of a "cysteine switch" that
contains a conserved cysteine residue which interacts with the zinc
in the active site and prevents binding and cleavage of the
substrate keeping the enzyme in an inactive form. In the majority
of the MMPs, the cysteine residue is in the conserved sequence
PRCGxPD. Some MMPs have a prohormone convertase cleavage site
(Furin-like) as part of this domain which when cleaved activates
the enzyme. MMP-23A and MMP-23B include a transmembrane segment in
this domain (PMID 10945999).
[1234] The MMPs can be subdivided in different ways. Use of
bioinformatic methods to compare the primary sequences of the MMPs
suggests the following evolutionary groupings of the MMPs: MMP-19;
MMPs 11, 14, 15, 16 and 17; MMP-2 and MMP-9; all the other
MMPs.
[1235] Analysis of the catalytic domains in isolation suggests that
the catalytic domains evolved further once the major groups had
differentiated, as is also indicated by the substrate specificities
of the enzymes. The most commonly used groupings (by researchers in
MMP biology) are based partly on historical assessment of the
substrate specificity of the MMP and partly on the cellular
localization of the MMP. These groups are the collagenases, the
gelatinases, the stromelysins, and the membrane type MMPs
(MT-MMPs). It is becoming increasingly clear that these divisions
are somewhat artificial as there are a number of MMPs that do not
fit into any of the traditional groups.
[1236] The collagenases are capable of degrading triple helical
fibrillar collagens into distinctive 3/4 and 1/4 fragments. These
collagens are the major components of bone and cartilage, and MMPs
are the only known mammalian enzymes capable of degrading them.
Traditionally, the collagenases are: MMP-1 (Interstitial
collagenase), MMP-8 (Neutrophil collagenase), MMP-13 (Collagenase
3), MMP-18 (Collagenase 4), MMP-14 (MT1-MMP) has also been shown to
cleave fibrillar collagen, and more controversially there is
evidence that MMP-2 is capable of collagenolysis.
[1237] The stromelysins display a broad ability to cleave ECM
proteins but are unable to cleave the triple-helical fibrillar
collagens. The three canonical members of this group are: MMP-3
(Stromelysin 1), MMP-10 (Stromelysin 2), and MMP-11 (Stromelysin
3). MMP-11 shows more similarity to the MT-MMPs, is
convertase-activatable and is secreted therefore usually associated
to convertase-activatable MMPs.
[1238] The main substrates of gelatinases are type IV collagen and
gelatin, and these enzymes are distinguished by the presence of an
additional domain inserted into the catalytic domain. This
gelatin-binding region is positioned immediately before the zinc
binding motif, and forms a separate folding unit which does not
disrupt the structure of the catalytic domain. The two members of
this sub-group are: MMP-2 (72 kDa gelatinase, gelatinase-A) and
MMP-9 (92 kDa gelatinase, gelatinase-B).
[1239] The secreted MMPs include MMP-11 (Stromelysin 3), MMP-21
(X-MMP), and MMP-28 (Epilysin).
[1240] The membrane-bound MMPs include: the type-II transmembrane
cysteine array MMP-23, the glycosyl phosphatidylinositol-attached
MMPs 17 and 25 (MT4-MMP and MT6-MMP respectively), and the type-I
transmembrane MMPs 14, 15, 16, 24 (MT1-MMP, MT2-MMP, MT3-MMP, and
MT5-MMP respectively).
[1241] All 6 MT-MMPs have a furin cleavage site in the pro-peptide,
which is a feature also shared by MMP-11.
[1242] Other MMPs include MMP-12 (Macrophage metalloelastase),
MMP-19 (RASI-1, occasionally referred to as stromelysin-4),
Enamelysin (MMP-20), and MMP-27 (MMP-22, C-MMP), MMP-23A (CA-MMP),
and MMP-23B.
Pharmaceutical Compositions and Administration
[1243] Once an imaging agent has been prepared or obtained, it may
be combined with one or more pharmaceutically acceptable excipients
to form a pharmaceutical composition that is suitable for
administering to a subject, including a human. As would be
appreciated by one of skill in this art, the excipients may be
chosen, for example, based on the route of administration as
described below, the agent being delivered, time course of delivery
of the agent, and/or the health/condition of the subject.
[1244] Pharmaceutical compositions of the present invention and for
use in accordance with the present invention may include a
pharmaceutically acceptable excipient or carrier. As used herein,
the term "pharmaceutically acceptable excipient" or
"pharmaceutically acceptable carrier" means a non-toxic, inert
solid, semi-solid or liquid filler, diluent, encapsulating material
or formulation auxiliary of any type. Some examples of materials
which can serve as pharmaceutically acceptable carriers are sugars
such as lactose, glucose, and sucrose; starches such as corn starch
and potato starch; cellulose and its derivatives such as sodium
carboxymethyl cellulose, ethyl cellulose, and cellulose acetate;
powdered tragacanth; malt; gelatin; talc; excipients such as cocoa
butter and suppository waxes; oils such as peanut oil, cottonseed
oil; safflower oil; sesame oil; olive oil; corn oil and soybean
oil; glycols such as propylene glycol; esters such as ethyl oleate
and ethyl laurate; agar; detergents such as Tween 80; buffering
agents such as magnesium hydroxide and aluminum hydroxide; alginic
acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl
alcohol; and phosphate buffer solutions, as well as other non-toxic
compatible lubricants such as sodium lauryl sulfate and magnesium
stearate, as well as coloring agents, releasing agents, coating
agents, sweetening, flavoring and perfuming agents, preservatives
and antioxidants can also be present in the composition, according
to the judgment of the formulator.
[1245] The pharmaceutical compositions of this invention can be
administered to humans and/or to animals parenterally such as
intravenously, intranasally (via a nasal spray), and
intraperitoneally. The mode of administration will vary depending
on the intended use, as is well known in the art. Alternatively,
formulations of the present invention may be administered
parenterally as injections (intravenous, intramuscular, or
subcutaneous). These formulations may be prepared by conventional
means, and, if desired, the subject compositions may be mixed with
any conventional additive.
[1246] Injectable preparations, for example, sterile injectable
aqueous or oleaginous suspensions may be formulated according to
the known art using suitable dispersing or wetting agents and
suspending agents. The sterile injectable preparation may also be a
sterile injectable solution, suspension, or emulsion in a nontoxic
parenterally acceptable diluent or solvent, for example, as a
solution in 1,3-butanediol. Among the acceptable vehicles and
solvents that may be employed are water, Ringer's solution, U.S.P.
and isotonic sodium chloride solution. In addition, sterile, fixed
oils are conventionally employed as a solvent or suspending medium.
For this purpose any bland fixed oil can be employed including
synthetic mono- or diglycerides. In addition, fatty acids such as
oleic acid are used in the preparation of injectables.
[1247] The injectable formulations can be sterilized, for example,
by filtration through a bacteria-retaining filter, or by
incorporating sterilizing agents in the form of sterile solid
compositions which can be dissolved or dispersed in sterile water
or other sterile injectable medium prior to use.
[1248] The imaging agents of the invention may be provided in any
suitable form, for example, in a pharmaceutically acceptable form.
In some cases, the imaging agent is included in a pharmaceutically
acceptable composition. In some embodiments, the imaging agent is
provided as a composition comprising ethanol, sodium ascorbate, and
water. In some cases, the composition comprises less than 20 weight
% ethanol, less than 15 weight % ethanol, less than 10 weight %
ethanol, less than 8 weight % ethanol, less than 6 weight %
ethanol, less than 5 weight % ethanol, less than 4 weight %
ethanol, less than 3 weight % ethanol, or less ethanol. In some
cases, the composition comprises less than 100 mg/mL, less than 75
mg/mL, less than 60 mg/mL, less than 50 mg/mL, less than 40 mg/mL,
less than 30 mg/mL, or less sodium ascorbate in water. In a
particular non-limiting embodiment, the imaging agent is provided
as a solution in water comprising less than 4% ethanol and less
than 50 mg/mL sodium ascorbate in water.
[1249] The imaging agent composition for injection may be prepared
in an injection syringe. The imaging agent may be prepared by a
radiopharmacy (e.g., using the methods described herein) and
provided to a health-care professional for administration. In some
aspects of the invention, the imaging agent is provided, for
example, in a syringe or other container, with .ltoreq.50 mg/mL
sodium ascorbate in water, .ltoreq.4 wt % ethanol, and about 1 to
14 mCi of the imaging agent. In some aspects of the invention, the
imaging agent is provided in a container such as a vial, bottle, or
syringe, and may be transferred, as necessary, into a suitable
container, such as a syringe for administration.
[1250] Syringes that include an adsorbent plunger tip may result in
10 to 25% of the imaging agent activity remaining in the syringe
after injection. Syringes lacking an adsorbent plunger tip may be
used, such as a 3 or 5 mL NORM-JECT (Henke Sass Wolf, Dudley,
Mass.) or other equivalent syringe lacking an adsorbent plunger
tip. Reduction of adsorption in the syringe can increase the amount
of the imaging agent that is transferred from the syringe and
administered to the subject in methods of the invention. A syringe
used in methods of the invention may comprise the imaging agent,
and be a non-adsorbing, or reduced adsorbent syringe. In some
embodiments a non-adsorbent or reduced-adsorbent syringe is a
syringe that has been coated or treated to reduce the imaging agent
adsorption. In some embodiments, a non-adsorbent or
reduced-adsorbent syringe is a syringe that lacks an adsorbent
plunger tip. In some embodiments, a syringe used in conjunction
with the invention adsorbs less than 20%, 19%, 18%, 17%, 16%, 15%,
14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or
0.5% of the imaging agent it contains. In certain aspects of the
invention, a syringe that contains the imaging agent does not
include a rubber or latex tip on the plunger. In some cases a
syringe used in methods of the invention, includes a plunger that
adsorbs less than 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%,
10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or 0.5% of the imaging
agent that the syringe contains. A syringe of the invention may
also comprise sodium ascorbate, ethanol, and water, and certain
embodiments of the invention include a syringe containing the
imaging agent in a solution comprising less than 4% ethanol and
less than 50 mg/mL sodium ascorbate in water. A syringe of the
invention may be a syringe that is latex free, rubber free, and/or
lubricant free. A syringe of the invention may contain the imaging
agent in an amount between about 1.5 and about 14 mCi. A syringe of
the invention may contain about 20 mCi or less of the imaging
agent.
[1251] Components of a composition comprising the imaging agent may
be selected depending on the mode of administration to the subject.
Various modes of administration that effectively deliver imaging
agents of the invention to a desired tissue, cell, organ, or bodily
fluid will be known to one of ordinary skill in the art. In some
embodiments, the imaging agent is administered intravenously (e.g.,
intravenous bolus injection) using methods known to those of
ordinary skill in the art. As used herein, a dose that is
"administered to a subject" means an amount of the imaging agent,
e.g. the imaging agent that enters the body of the subject. In some
embodiments, due to factors such as partial retention of imaging
agent such as the imaging agent in a syringe, tubing, needles,
catheter, or other equipment used to administer the imaging agent
to a subject, the amount of an imaging agent such as the imaging
agent that is measured or determined to be in the a syringe or
other equipment prepared for administration may be more than the
amount in the dose that is administered to the subject. In some
embodiments, an injection of an imaging agent is followed by a
flushing injection of normal saline, into the subject, using the
same tubing, needle, port, etc., used for administration of the
imaging agent. Flushing may be performed immediately following
administration of the imaging agent, or up to 1 min, 2 min, 3 min,
5 min, or more, after the administration. The volume of saline or
other agent for flushing may be up to 5 ml, 6 ml, 7 ml, 8 ml, 9 ml,
10 ml, 15 ml, 20 ml, or more. As will be understood by those of
ordinary skill in the art, in embodiments where the imaging agent
is administered using a syringe or other container, the true amount
of the imaging agent administered to the subject may be corrected
for any the imaging agent that remains in the container. For
example, the amount of radioactivity remaining in the container,
and tubing and needle or delivery instrument that carried the
imaging agent from the container and into the subject can be
determined after the imaging agent has been administered to the
subject and the difference between the starting amount of
radioactivity and the amount remaining after administration
indicates the amount that was delivered into the subject. In some
cases, the container or injection device (e.g., catheter, syringe)
may be rinsed with a solution (e.g., saline solution) following
administration of the imaging agent.
[1252] In some embodiments of the invention, the total amount of
the imaging agent administered to a subject over a given period of
time, e.g., in one session, is less than or equal to about 50 mCi,
less than or equal to 40 mCi, less than or equal to 30 mCi, less
than or equal to 20 mCi, less than or equal to 18 mCi, less than or
equal to 16 mCi, less than or equal to 15 mCi, less than or equal
to 14 mCi, less than or equal to 13 mCi, less than or equal to 12
mCi, less than or equal to 10 mCi, less than or equal to 8 mCi,
less than or equal to 6 mCi, less than or equal to 4 mCi, less than
or equal to 2 mCi, less than or equal to 1 mCi, less than or equal
to 0.5 mCi. The total amount administered may be determined based
on a single dose or multiple doses administered to a subject within
a given time period of up to 1 minute, 10 minutes, 30 minutes, 1
hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 hours, or more.
[1253] Based on radiation dose studies, the desirable maximum dose
administered to a subject may be based on determining the amount of
the imaging agent which limits the radiation dose to about 5 rem to
the critical organ and/or about 1 rem effective dose (ED) or lower,
as will be understood by those of ordinary skill in the art. In a
particular embodiment, the desirable maximum dose or total amount
of the imaging agent administered is less than or equal to about 25
mCi, or less than or equal to about 14 mCi over a period of time of
up to 30 min, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48
hours, or more. In some embodiments, the maximum dose of the
imaging agent administered to a subject may be less than 3.5 .mu.g
per 50 kg of body weight per day. That is, in some embodiments of
the invention, the maximum dose of the imaging agent administered
to a subject may be less than about 0.07 .mu.g of the imaging agent
per kg of body weight per day.
Abbreviations
[1254] The following abbreviations are used herein: [1255] Acm
acetamidomethyl [1256] b-Ala, beta-Ala or bAla 3-aminopropionic
acid [1257] ATA 2-aminothiazole-5-acetic acid or
2-aminothiazole-5-acetyl group [1258] Boc t-butyloxycarbonyl [1259]
CBZ, Cbz or Z Carbobenzyloxy [1260] Cit citrulline [1261] Dap
2,3-diaminopropionic acid [1262] DCC dicyclohexylcarbodiimide
[1263] DIEA diisopropylethylamine [1264] DMAP
4-dimethylaminopyridine [1265] EOE ethoxyethyl [1266] HBTU
2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium
hexafluorophosphate [1267] hynic boc-hydrazinonicotinyl group or
2-[[[5-[carbonyl]-2-pyridinyl]hydrazono]methyl]-benzenesulfonic
acid, [1268] NMeArg or MeArg a-N-methyl arginine [1269] NMeAsp
a-N-methyl aspartic acid [1270] NMM N-methylmorpholine [1271] OcHex
O-cyclohexyl [1272] OBzl O-benzyl [1273] oSu O-succinimidyl [1274]
TBTU 2-(1H-Benzotriazol-1-yl)-1,1,3,3-tetramethyluronium
tetrafluoroborate [1275] THF tetrahydrofuranyl [1276] THP
tetrahydropyranyl [1277] Tos tosyl [1278] Tr trityl
[1279] The following conventional three-letter amino acid
abbreviations are used herein; the conventional one-letter amino
acid abbreviations are not used herein:
[1280] Ala=alanine Arg=arginine Asn=asparagine Asp=aspartic acid
Cys=cysteine Gln=glutamine Glu=glutamic acid Gly=glycine
His=histidine Ile=isoleucine Leu=leucine Lys=lysine Met=methionine
Nle=norleucine Orn=ornithine Phe=phenylalanine Phg=phenylglycine
Pro=proline Sar=sarcosine Ser=serine Thr=threonine Trp=tryptophan
Tyr=tyrosine Val=valine.
EXAMPLES
Example 1
[1281] We have established the feasibility of in vivo imaging of
MMP activation in pigs (Sahul et al. Circ Cardiovasc Imaging 2011,
4:381-391) and dogs (Liu et al. J Nucl Med 2011, 52(3):453-60)
post-MI. The data derived in pigs involved surgical occlusion of
two marginal branches of the left circumflex artery and resulted in
regional activation of MMPs in the inferolateral wall. (Sahul et
al. Circ Cardiovasc Imaging 2011, 4:381-391). This surgical model
caused significant activation of MMPs in the surgical wound
adjacent to both the atria and ventricles of heart, complicating in
vivo imaging. The studies in dogs employed percutaneous balloon
occlusion of left anterior descending artery, avoided the surgical
intervention, and resulted in improved image quality. In these
recently published porcine studies with serial SPECT/CT imaging, we
demonstrated focal uptake of the MMP-targeted agent
.sup.99mTc-RP805 within the infarcted lateral wall, which peaked at
.about.2 weeks post injury, and remained elevated at 4 weeks post
occlusion. Early MMP activity at 1 week post-MI predicted late post
MI ventricular remodeling (FIG. 1).
Example 2
[1282] Heart failure after MI leads to atrial remodeling and
fibrosis, thereby increasing vulnerability to AF. The role of
atrial MMP activation has not been well studied. We hypothesized
that atrial structural remodeling and fibrillation vulnerability
occurring early after MI can be noninvasively assessed using
targeted molecular imaging of MMP activation.
[1283] Methods:
[1284] In vivo and ex vivo SPECT/CT images were obtained in control
pigs (n=7) and in pigs 10 days (n=6) or 4 weeks (n=6) after
surgical induction of MI. MI was induced by surgical ligation of
two marginal branches of the left circumflex coronary artery.
Animals were injected intravenously with a .sup.99mTc-labeled
radiotracer (.sup.99mTc-RP805) targeted to activated MMPs. Hybrid
64-slice SPECT/CT scans were acquired at 2 hours after injection of
a .sup.99mTc-RP805. X-ray CT imaging with contrast was performed to
define coronary anatomy and cardiac chambers.
After sacrifice, hearts were excised and cast in alginate for ex
vivo SPECT/CT imaging. Myocardial .sup.99mTc-RP805 retention in the
atria and the ventricles was quantified by gamma well counting
after sacrifice. AF vulnerability was assessed in subsets of
control pigs (n=5) and in pigs 4 weeks post-MI (n=4) using atrial
burst pacing for 10 seconds with cycle lengths ranging from 300 to
180 ms.
[1285] Results:
[1286] In vivo and ex vivo SPECT/CT imaging demonstrated increased
.sup.99mTc-RP805 retention in the MI region and both atria compared
to control pigs. At 10 days post-MI, .sup.99mTc-RP805 retention (%
injected dose/gram) was increased .about.6-fold in the MI region
and .about.3-fold in the left atrium (LA) compared to control
regions (p<0.01 each, FIG. 2, bottom panel). At 4 weeks post-MI,
99Tc-RP805 retention was increased .about.4-fold in the MI region
and .about.2-fold in the LA compared to control regions (p<0.01
each, FIG. 2, bottom panel). AF was inducible in 4 of 4 pigs at 4
weeks post-MI and 0 of 5 controls (p<0.01). Representative in
vivo and ex vivo images at 10 days post-MI are shown in FIG. 2, top
and middle panels.
[1287] Table 1 shows that AF burden is significantly increased at
four weeks post-MI in our post-MI HF model. Following 10 seconds of
burst pacing at cycle lengths ranging from 260-180 ms, the duration
of AF in post-MI animals was 2.0.+-.1.8 minutes. No AF could be
induced in the control group despite pacing down to a cycle length
of 180 ms.
TABLE-US-00002 TABLE 1 AF duration Burst Cycle Length (sec) (ms)
p-value Control (n = 3) 0.0 215 .+-. 30 4-week HF (n = 4) 2.0 .+-.
1.8 <180 0.03
[1288] Conclusions:
[1289] MMP-targeted SPECT/CT imaging provides a valuable
noninvasive approach for assessment of atrial remodeling and allows
early identification of arrhythmogenic substrates prior to the
onset of irreversible fibrosis. In vivo imaging of MMP activation
has significant clinical implications regarding risk stratification
and directing pharmacological and interventional treatments of
AF.
Example 3
[1290] A cohort of patients post cardioversion are administered an
effective amount of the imaging agent of the invention, images of
each patient's left atrium are obtained and the uptake of the
imaging agent is quantified. A cut point for imaging agent uptake
is then established such that the cut point separates the cohort
into 2 populations; those above the cut point have recurrent AF
while those below the cut point do not.
[1291] Based on the cut point levels determined above, future
patients are then tested for MMP levels using the methods of the
invention and those demonstrating above cut point levels are
treated using one or more of the therapies described herein and
known in the art for AF, including but not limited to
pharmacological rate control therapy, pharmacological rhythm
control therapy, ablation, and/or implantable pacer.
Example 4
[1292] A cohort of patients with a recent history of myocardial
infarction are administered an effective amount of the imaging
agent of the invention, images of each patient's left atrium are
obtained and the uptake of the imaging agent is quantified. The
patients also undergo a resting flurpiridaz F 18 myocardial
perfusion study and the summed rest score determined for each
patient. Logistic regression analysis is performed to produce an
equation expressing the likelihood of future AF as a function of
summed rest score and quantified imaging agent uptake.
Example 5
Part A
Preparation of Imaging Agent 1
##STR00090##
[1294]
Methyl(3S,7S,6R)-4-aza-7-[(tert-butyl)oxycarbonyl]-6-(2-methylpropy-
l)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-triene-3-carboxylat-
e was prepared according to the method of Xue, et al. (J. Med.
Chem. 2001, 44, 2636-2660) then transformed into Imaging Agent 1
through the convergent assembly of
O-(tetrahydro-2H-pyran-2-yl)hydroxylamine and
3-fluoropropan-1-amine using standard protecting group strategy
(Wuts, P. G. M.; Greene, T. W. The Role of Protective Groups in
Organic Synthesis. In Greene's Protective Group in Organic
Synthesis, Fourth Edition; John Wiley & Sons, Inc.: Hoboken,
N.J., 2007; pp 1-15) and peptide coupling chemical methods (Tsuda,
Y; Okada, Y. Solution-Phase Peptide Synthesis. In Amino Acids,
Peptides and Proteins in Organic Chemistry: Building Blocks,
Catalysis and Coupling Chemistry, Volume 3; Hughes, A. B. Ed.;
Wiley-VCH Verlag GmbH & Co. KgaA: Weinheim, Germany, 2010;
201-251) commonly known to those skilled in the art.
Part B
Preparation of Imaging Agent 2
##STR00091##
[1296]
Methyl(3S,7S,6R)-4-aza-7-[(tert-butyl)oxycarbonyl]-6-(2-methylpropy-
l)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-triene-3-carboxylat-
e was prepared according to the method of Xue, et al. (J. Med.
Chem. 2001, 44, 2636-2660) then transformed into Imaging Agent 2
through the convergent assembly of
O-(tetrahydro-2H-pyran-2-yl)hydroxylamine and 6-aminohexan-1-ol
using standard protecting group strategy, peptide coupling, and
fluorination chemical methods commonly known to those skilled in
the art.
Part C
Preparation of Imaging Agent 3
##STR00092##
[1298] Methyl(3S,7S,
6R)-4-aza-7-[(tert-butyl)oxycarbonyl]-6-(2-methylpropyl)-11-oxa-5-oxobicy-
clo[10.2.2]hexadeca-1(15),12(16),13-triene-3-carboxylate was
prepared according to the method of Xue, et al. (J. Med. Chem.
2001, 44, 2636-2660) then transformed into Imaging Agent 3 through
the convergent assembly of
O-(tetrahydro-2H-pyran-2-yl)hydroxylamine,
2-((tert-butoxycarbonyl)amino)acetic acid and
3-fluoropropan-1-amine using standard protecting group strategy and
peptide coupling chemical methods commonly known to those skilled
in the art.
Part D
Preparation of Imaging Agent 4
##STR00093##
[1300]
Methyl(3S,7S,6R)-4-aza-7-[(tert-butyl)oxycarbonyl]-6-(2-methylpropy-
l)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-triene-3-carboxylat-
e was prepared according to the method of Xue, et al. (J. Med.
Chem. 2001, 44, 2636-2660) then transformed into Imaging Agent 4
through the convergent assembly of
O-(tetrahydro-2H-pyran-2-yl)hydroxylamine,
2-((tert-butoxycarbonyl)amino)acetic acid and 5-aminopentan-1-ol
using standard protecting group strategy, peptide coupling, and
fluorination chemical methods commonly known to those skilled in
the art.
Part E
Preparation of Imaging Agent 5
##STR00094##
[1302]
Methyl(3S,7S,6R)-4-aza-7-[(tert-butyl)oxycarbonyl]-6-(2-methylpropy-
l)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1(15),12(16),13-triene-3-carboxylat-
e was prepared according to the method of Xue, et al. (J. Med.
Chem. 2001, 44, 2636-2660) then transformed into Imaging Agent 5
through the convergent assembly of
O-(tetrahydro-2H-pyran-2-yl)hydroxylamine and
(4-fluorophenyl)methanamine using standard protecting group
strategy and peptide coupling chemical methods commonly known to
those skilled in the art.
Part F Preparation of Imaging Agent 6
##STR00095##
[1304]
Methyl(3S,7S,6R)-4-aza-7-[(tert-butyl)oxycarbonyl]-6-(2-methylpropy-
l)-11-oxa-5-oxobicyclo[10.2.2]hexadeca-1
(15),12(16),13-triene-3-carboxylate was prepared according to the
method of Xue, et al. (J. Med. Chem. 2001, 44, 2636-2660) then
transformed into Imaging Agent 6 through the convergent assembly of
O-(tetrahydro-2H-pyran-2-yl)hydroxylamine, and
(4-(3-fluoropropoxy)phenyl)methanamine using standard protecting
group strategy and peptide coupling chemical methods commonly known
to those skilled in the art. (4-(3-Fluoropropoxy)phenyl)methanamine
was prepared from 4-hydroxybenzonitrile and
1-bromo-3-fluoropropane.
Example 6
In-Vitro MMP Inhibition Assay
[1305] Individual inhibitors were dissolved in TCN buffer (50 mM
Tris-HCl, 10 mM CaCl.sub.2, 150 mM NaCl.sub.2, 0.05% Brij 35 at pH
7.5) at appropriate dilutions then added to the wells of a
microtiter plate (10 .mu.L/well) in triplicate. Each well of test
agent (and appropriate control wells) was then treated with
activated MMP-2 or 9 (10 .mu.L of a 40 nM solution in 50 mM Hepes,
10 mM CaCl.sub.2, 1% Brij 35 at pH 7.5; R&D Systems) followed
by 30 .mu.L TCN buffer and 150 .mu.L the fluorogenic peptide
substrate (Mca-PLGL-Dpa-AR-NH.sub.2; R&D Systems). The
resulting mixtures were incubated 1 h at 27.degree. C. then
analyzed using a FL600 fluorescent plate reader (excitation=310/20;
emission=420/50; optics=bottom; sensitivity=225) and KC4
Software.
TABLE-US-00003 TABLE 2 MMP inhibition data Imaging Agent MMP-2
MMP-9 1 5.19 3.54 2 31.6 20.2 3 4.61 2.59 4 15.4 15.3 5 0.58 0.74 6
2.74 2.98 RP805 6.50 7.40
Example 7
[1306] FIG. 3 shows representative transaxial slices from ex vivo
SPECT images of a control pig heart, and hearts from pigs at 1 and
2 weeks post-MI. the top row of each image set are targeted
.sup.99mTc-RP805 images (linear grey scale) matched with
corresponding high resolution CT images (gray scale, below). Hearts
were filled with alginate mixed with CT contrast to define right
and left ventricles (RV & LV) and atria (RA & LA). Uniform
uptake is seen in the control heart. Infarcted hearts demonstrate
focal .sup.99mTc-RP805 in both the infarct region and atria.
[1307] FIG. 4 shows results from quantitative analysis of ex vivo
.sup.99mTc-RP805 SPECT images from a pig heart at 4 weeks post-MI.
A. Uncorrected SPECT images, B. SPECT images with resolution
recovery, C. SPECT data reconstructed with partial volume
correction (PVC), D. Grey scale-coded .sup.99mTc-RP805 activity for
8 radial sectors per slice from gamma well counter, E. Postmortem
images of heart demonstrating dense inferolateral scar and marked
wall thinning, F. Correlation between measured regional myocardial
well-counter activity and SPECT derived activity with PVC.
[1308] FIG. 5 shows a plot of the MMP total activity per unit time,
according to some embodiments.
[1309] FIG. 6 shows a plot of the percent area collagen for the
left and right regions of the heart. Regional changes in matrix
structure could be detected by 1 week post-MI which were
significant by 4 weeks post-MI (Kruskal-Wallis, p<0.05). In the
inset: PSR imaging revealed matrix disruption and discontinuity
between atrial myocytes which was progressive with time
post-MI.
Example 8
[1310] Calcific aortic valve disease (CAVD) is common among the
elderly population. Inflammation and matrix remodeling play a
central role in the progression of CAVD to symptomatic aortic
stenosis. Matrix metalloproteinases (MMPs) are upregulated in CAVD.
In vivo imaging of MMP activation may lead to prospective
identification of aortic valves that are at high risk for
developing stenosis and help track the effect of potential novel
therapeutic interventions. This example illustrates use of an
MMP-targeted agent for both in vivo imaging and definition of
temporal patterns of MMP activation in CAVD.
[1311] Methods and Results:
[1312] ApoE.sup.-/- mice were fed a high fat diet (HFD) for up to 9
months. Histological analysis of the aortic valve showed
considerable thickening of valve leaflets over time. M mode
echocardiography demonstrated a reduction in leaflet separation
from 3 months to 9 months. Non-contrast high resolution CT
established the presence of aortic valve calcification after 9
months of HFD. MMP-targeted microSPECT imaging using
.sup.99mTc-RP805, a tracer with specificity for activated MMPs,
followed by CT angiography showed considerable tracer uptake (in
counts per voxel per MBq injected dose) in the aortic valve area at
3, 6 and 9 months. Uptake was maximal after 6 months of HFD (3 m:
0.047.+-.0.002, n=2, 6 m: 0.102.+-.0.013, n=4, 9 m: 0.064.+-.0.004,
n=4). Tracer uptake in the aortic valve area was confirmed
following ex vivo planar imaging.
[1313] Conclusion:
[1314] MMP-targeted microSPECT/CT imaging can detect aortic valve
biology in CAVD in vivo. In this model, protease activation in the
aortic valve is maximal at 6 months and declines with progression
of CAVD.
[1315] FIG. 7 shows .sup.99mTc-RP805 in vivo microSPECT/CT imaging
(left) of MMP activation in an ApoE-/- mouse fed a Western diet for
9 months. Tracer uptake in the aortic valve area is indicated by
the arrows. Uptake in the aortic valve was confirmed by ex vivo
planar imaging (right) of the explanted heart and aorta.
[1316] FIG. 8 shows in vivo uptake of .sup.99mTc-RP805 in the
aortic valve over time in ApoE.sup.-/- mice fed a Western diet.
[1317] FIG. 9 shows ex vivo uptake of .sup.99mTc-RP805 in the
aortic valve over time in ApoE.sup.-/- mice fed a Western diet.
[1318] FIG. 10 shows autoradiography of the explanted aorta from an
ApoE-/- mouse fed a Western diet for three months. Arrows indicate
uptake of .sup.111In-RP782 in the aortic valve area.
[1319] FIG. 11 shows H&E staining of the aortic valve in
ApoE-/- mice fed a Western diet for 4 (left) and 9 months (right)
demonstrating marked remodeling of valve leaflets over time.
[1320] FIG. 12 shows immunostaining of F4-80 (dark grey) in the
aortic valve from an ApoE-/- mouse fed a Western diet for 6 months
demonstrating considerable macrophage infiltration.
[1321] FIG. 13 shows plots of aortic valve GAPDH-normalized CD68
(top) and MMP-12 (bottom) mRNA expression quantified by real time
RT-PCR in wild type (WT) mice on normal chow and ApoE-/- mice fed a
Western diet for 3, 6 or 9 months.
[1322] FIG. 14 shows grey scale-coded non contrast CT images of an
ApoE-/- mouse fed a Western diet for 10 months demonstrating
calcification of the aortic valve. Arrows indicate the aortic valve
plane.
[1323] FIG. 15 shows M mode echocardiographic images of ApoE-/-
mice fed a Western diet demonstrating normal systolic separation of
aortic valve cusps after 3 months on diet (left) and reduced
separation after 9 months (right).
Example 9
[1324] Atherosclerosis, a major cause of morbidity and mortality in
the US, is linked to hyperlipidemia. Pharmacologic treatment of
hyperlipidemia is a mainstay of modern treatment for
atherosclerotic diseases and is believed to be related at least in
part to "stabilizing" effects on plaque biology. This example
investigates the effect of anti-lipid therapies on plaque biology
through serial imaging of matrix metalloproteinase (MMP) activation
in vivo.
[1325] Methods:
[1326] ApoE.sup.-/- mice were fed a high fat diet (HFD) for 2
months to induce atherosclerosis. After two months, the mice were
randomly assigned to one of 4 groups: HFD, HFD plus simvastatin
(Sim), HFD plus fenofibrate (Fen) and high fat withdrawal (HFW).
MicroSPECT/CT imaging using .sup.99mTc-RP805, a tracer with
specificity for activated MMPs was performed after one week and 4
weeks.
[1327] Results:
[1328] Withdrawal of the HFD significantly reduced total
cholesterol levels at 1 week (1845.1.+-.41.9 to 492.7.+-.19.4
mg/dL, p=0.001). Neither simvastatin nor fenofibrate had a
significant effect on total cholesterol level compared to animals
on HFD at one week, but both significantly reduced cholesterol
levels by 4 weeks. At 1 week, there was no significant difference
in uptake of .sup.99mTc-RP805 in the aortic arch between different
experimental groups. Uptake (in counts per voxel per mCi injected
dose) at 4 weeks however, was significantly higher in the HFD group
compared to other three groups (HFD: 4.99.+-.0.27, n=5, Sim:
3.17.+-.0.52, n=6, p<0.02, Fen: 2.23.+-.0.28, n=7, p<0.001,
HFW: 1.77.+-.0.22, n=5, p<0.001). .sup.99mTc-RP805 uptake in the
aortic arch significantly increased from 1 week to 4 weeks in
animals on HFD (mean uptake 2.84.+-.0.47 versus 4.99.+-.0.27,
p<0.003), but did not occur in the other experimental
groups.
[1329] Conclusions:
[1330] MMP-targeted molecular imaging demonstrates an effect of
anti-lipid therapies on plaque biology at 4 weeks, but not at 1
week after initiation of therapy.
Example 10
Tissue Biodistribution
[1331] Oncomice.RTM., obtained through an in-house breeding
program, were anesthetized intramuscularly with 0.1 mL of
ketamine/acepromazine (1.8 mL saline, 1.0 mL ketamine, and 0.2 mL
acepromazine) prior to dosing and tissue sampling. Individual mice
were then injected via the tail vein with an imaging agent of the
present invention (0.5-2.0 mCi/kg in 0.1 mL). Mice were euthanized
and biodistribution performed at 1 h post-injection. Selected
tissues were removed, weighed, and counted on a gamma counter.
Results are expressed as the percentage of injected dose per gram
tissue (mean.+-.SEM; Table 3).
TABLE-US-00004 TABLE 3 Summary of imaging agent distribution in the
Oncomonse .RTM. Imaging Agent Distribution (% ID/g) tissue 2 4 6
blood 1.07 .+-. 0.060 0.41 .+-. 0.099 0.88 .+-. 0.061 heart 0.95
.+-. 0.065 0.36 .+-. 0.064 0.69 .+-. 0.073 lung 0.97 .+-. 0.121
0.45 .+-. 0.071 1.69 .+-. 0.382 liver 13.1 .+-. 2.17 23.6 .+-. 5.19
11.3 .+-. 1.73 spleen 0.69 .+-. 0.085 0.34 .+-. 0.057 0.81 .+-.
0.021 kidney 20.6 .+-. 3.25 14.8 .+-. 1.79 6.66 .+-. 1.46 bone 2.02
.+-. 0.320 1.28 .+-. 0.200 2.86 .+-. 0.124 muscle 0.50 .+-. 0.073
0.17 .+-. 0.043 0.44 .+-. 0.049 urine 71.8 7.67 .+-. 5.00 7.21 .+-.
6.71 tumor 0.95 .+-. 0.103 1.12 .+-. 0.204 0.73 .+-. 0.026
EQUIVALENTS
[1332] Unless defined otherwise, all technical and scientific terms
used herein have the same meanings as commonly understood by one of
skill in the art to which the disclosed methods and compositions
belong.
[1333] Throughout this application, various publications are
referenced. The disclosures of these publications in their
entireties are hereby incorporated by reference into this
application in order to more fully describe the state of the art to
which this pertains. The references disclosed are also individually
and specifically incorporated by reference herein for the material
contained in them that is discussed in the sentence in which the
reference is relied upon.
[1334] It must be noted that as used herein and in the appended
claims, the singular forms "a," "an," and "the" include plural
reference unless the context clearly dictates otherwise. Thus, for
example, reference to "an MMP" includes a plurality of such MMPs,
reference to "the MMP" is a reference to one or more MMP and
equivalents thereof known to those skilled in the art, and so
forth.
[1335] "Optional" or "optionally" means that the subsequently
described event, circumstance, or material may or may not occur or
be present, and that the description includes instances where the
event, circumstance, or material occurs or is present and instances
where it does not occur or is not present.
[1336] Throughout the description and claims of this specification,
the word "comprise" and variations of the word, such as
"comprising" and "comprises," means "including but not limited to"
and is not intended to exclude, for example, other additives,
components, integers or steps.
* * * * *